Amplification: Consumables

Amplification Reagents and Plastics

Factors Impacting Gene Expression Analysis
RNA Isolation
RNA integrity, purity, and yield
■■

Genomic DNA contamination

■■

Inhibitors of cDNA synthesis and qPCR

■■

RNase and DNase contamination

■■

Reagents ­— Reverse Transcription

cDNA synthesis efficiency
■■

RNA protection
■■

Input RNA capacity

■■

Accurate representation of mRNA

■■

Reagents — ­ Real-Time qPCR

Detection sensitivity
■■

■■ Inhibitors in sample
■■ Reproducibility of thermal cycling conditions and instrument compatibility
PrimePCR™ Assays and Panels
Primer design
■■

Assay validation
■■

Pathway curation
■■

PCR Plastic Consumables

Instrument compatibility
■■

Optimum performance
■■

Automation friendly
■■
RNA Isolation
■■  its are designed and formulated to assist
K
in the isolation of highly pure and intact RNA
from different starting materials
■■  NA is compatible with a variety
R
of downstream applications
– Real-time qPCR
– Northern blotting
– Microarray analysis
– cDNA library construction
■■ DNase treatment ensures genomic DNA removal
 RNA Isolation ­

RNA Isolation
Aurum™ Total RNA Fatty and Fibrous Tissue Kit Aurum Total RNA 96 Kit
■■ PCR-ready RNA in less than 60 min ■■ High-throughput total RNA isolation in less than 60 min
■■ PureZOL™ efficiently lyses cells and tissues, deproteinates RNA, and ■■  igh yield of intact total RNA from a wide range of starting materials,
H
inactivates endogenous nucleases in a single step including cultured cells, bacteria, and yeast, as well as plant and animal tissues
■■  uanidine isothiocyanate and b-mercaptoethanol efficiently lyse samples
G
■■  igh yield of intact total RNA from difficult-to-disrupt samples, including
H
and quickly inactivate RNases
plant and animal tissues
■■  Nase-free reagents and plastic consumables ensure the integrity
R
■■ Well-suited for fungal samples that are rich in RNases of isolated RNA
■■  Nase-free reagents and plastic consumables ensure the integrity
R ■■ Kit includes DNase I for removal of genomic DNA contamination
of isolated RNA ■■ Compatible with Aurum vacuum manifold
■■ Kit includes DNase I for removal of genomic DNA contamination For more information, download or request bulletin 2919.
www.bio-rad.com/
■■ Easy-to-use spin or vacuum protocol rna-isolation

For more information, download or request bulletin 5282.

Aurum Total RNA Mini Kit

■■ PCR-ready RNA in less than 60 min
■■  uanidine isothiocyanate and b-mercaptoethanol efficiently lyse samples
G
and quickly inactivate RNases
■■  igh yield of intact total RNA from a wide range of starting materials, including
H
cultured cells, bacteria, and yeast, as well as plant and animal tissues
732-6800, 2 x 96-well preps
■■  Nase-free reagents and plastic consumables ensure the integrity
R Aurum Total RNA 96 Kit
of isolated RNA
■■ Kit includes DNase I for removal of genomic

DNA contamination
■■ Easy-to-use spin or vacuum protocol
For more information, download or request bulletin 2920.

732-6830, 50 preps 732-6820, 50 preps

Aurum Total RNA Fatty and Fibrous Tissue Kit Aurum Total RNA Mini Kit

© 2013 Bio-Rad Laboratories, Inc. Amplification Reagents and Plastics 5

 RNA Isolation ­

iScript™ RT-qPCR Sample Preparation Reagent PureZOL™ RNA Isolation Reagent

■■ Reagent stabilizes RNA and removes genomic DNA in less than 10 min ■■  ingle-solution format permits recovery of RNA from small quantities of
S
www.bio-rad.com/ tissues or cells, making it ideally suited for gene expression studies
iscript
■■ Suitable for adherent or suspension animal cells
■■  T-qPCR is directly enabled from cells without RNA purification when
R
■■  fficient RNA purification from cultured cells, yeast, viruses, and bacteria,
E
combined with an iScript reverse transcription kit and real-time supermix as well as plant and animal tissues
■■  eagent allows multiplex real-time detection of up to 4 targets from as few
R
■■  ureZOL efficiently lyses cells and tissues, deproteinates RNA, and
P
as 10 cells inactivates endogenous nucleases in a single step
■■ Ideal for rapid, high-throughput gene expression analysis
■■ Scalable starting sample amount

For more information, download or request bulletin 5736.

■■ Convenient isolation of RNA, DNA, and protein from the same sample

104 Aurum™ Vacuum Manifold

■■ Vacuum-mediated nucleic acid purification platform
■■  ersatile manifold format adaptable for 96-well plate or up to
V
10 3 18 spin columns
RFU

■■  anifold ensures fast, high-quality sample preparation while

M
maintaining the simplicity of vacuum processing
10 2
■■  nique vacuum regulator design allows for complete control of
U
negative pressure
0 10 20 30 40
Cycles
iScript RT-qPCR sample preparation reagent generates linear
results over varying input cell amounts. HeLa cells (125, 25, 5, and
1 cells/µl) were treated and analyzed for GAPDH expression levels
using iScript cDNA synthesis kit and iQ ™ SYBR ® Green supermix
on the CFX96™ real-time PCR detection system. RFU, relative
fluorescence units.

732-6890, 100 ml
PureZOL RNA Isolation Reagent

170-8899, 5 x 10 ml 732-6470, 1 unit

iScript RT-qPCR Sample Preparation Reagent Aurum Vacuum Manifold

Amplification Reagents and Plastics 6 Visit us on the Web at www.bio-rad.com.

 RNA Isolation ­
Selection Guide

RNA Isolation
Aurum™ Total RNA Kits PureZOL™ RNA
Mini Fatty and Fibrous Tissue 96 Isolation Reagent
Format Mini column Mini column 96-well plate Single solution
Filtration (vacuum or spin) Filtration (vacuum or spin) Filtration (vacuum or spin) organic extraction
Maximum starting
material amounts
Cultured cells 2 x 106 1 x 107 1 x 106 1 x 107
Bacterial cells 2.4 x 109 2.4 x 109 8 x 108 2.4 x 109
Yeast cells 3 x 107 3 x 107 2 x 107 3 x 107
Hard animal tissue 20 mg 100 mg — 100 mg
Soft to moderately 40 mg 100 mg — 100 mg
hard animal tissue
Plant tissue 40 mg 100 mg — 100 mg
Isolation method Silica membrane Lysis with PureZOL reagent, Silica membrane Organic extraction
purification on silica membrane
Number of preps 50 mini preps 50 mini preps 2 x 96-well plate 50 or 100 (1 ml/prep)
Number of washes 3 3 3 —
DNase I included* Yes Yes Yes No
DNase I digest time 15 min (animal tissue, 25 min) 15 min 10 min —
Total preparation time** <50–80 min (with DNase I digest) <50–80 min (with DNase I digest) <60 min (with DNase I digest) <60 min
Binding capacity >100 µg >100 µg >40 µg —
Elution volume 2 x 40 µl 2 x 40 µl 80 µl 30–100 µl
* Removal not required.
** Total preparation time will vary depending on the tissue or cell type and on which format is used (vacuum or spin).
For sample-specific yield information, please visit www.bio-rad.com/rna-isolation and click the RNA Isolation Selection Guide.

Ordering Information
Catalog # Description Catalog # Description
732-6830 Aurum Total RNA Fatty and Fibrous Tissue Kit 170-8899 iScript RT-qPCR Sample Preparation Reagent, 500 reactions
732-6870* Aurum Total RNA Fatty and Fibrous Tissue Module 732-6880 PureZOL RNA Isolation Reagent, 50 ml
732-6820  Aurum Total RNA Mini Kit 732-6890 PureZOL RNA Isolation Reagent, 100 ml
732-6800  Aurum Total RNA 96 Kit 732-6470 Aurum Vacuum Manifold
170-8898 iScript RT-qPCR Sample Preparation Reagent, 100 reactions

* Not provided with PureZOL RNA isolation reagent (see catalog #732-6890 or #732-6880 to order separately).

© 2013 Bio-Rad Laboratories, Inc. Amplification Reagents and Plastics 7

Reagents—
Reverse Transcription
■■ 
Formulated for efficient reverse transcription
across a broad linear dynamic range
■■  otent RNase A inhibitors protect RNA during
P
setup and reverse transcription
■■  lexible input RNA capacity to suit different
F
experimental needs
■■  ptimized for gene expression analysis using
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real-time qPCR
 Reagents
Reverse Transcription — iScript™ Kit Selector

Reduce Maximize data Fast and Select my One-step One-step

pipetting from single easy to use own primers RT-qPCR with RT-qPCR for
variability 20 μl reaction SYBR® Green probes

iScript reverse iScript advanced iScript cDNA iScript Select iScript™ one-step iScript one-step
transcription supermix cDNA synthesis kit synthesis kit cDNA synthesis kit RT-PCR kit RT-PCR kit for probes

Reagents
for RT-qPCR for RT-qPCR with SYBR® Green
Sensitivity

1 µg–100 fg 7.5 µg–100 fg 1 µg–100 fg 1 µg–1 pg 100 ng–1 pg 1 µg–1 pg

total RNA total RNA total RNA total RNA total RNA total RNA

iScript reverse iScript reverse iScript reverse iScript reverse iScript reverse
transcriptase transcriptase transcriptase transcriptase transcriptase
(for one-step RT-PCR) (for one-step RT-PCR)

5x iScript 5x iScript advanced 2x SYBR® Green

RT supermix 5x iScript reaction mix RT-PCR reaction mix 2x probes RT-PCR
Kit Contents

reaction mix 5x iScript reaction mix

(dNTPs, oligo[dT], (dNTPs, oligo[dT], (dNTPs and (dNTPs, iTaq™ DNA reaction mix (dNTPs,
random primers, (dNTPs, oligo[dT], random primers, and polymerase, iTaq DNA polymerase,
random primers, and buffer components) fluorescein,
buffer components, buffer components) and stabilizers)
and iScript reverse buffer components) SYBR® Green I dye,
transcriptase) and stabilizers)

Oligo(dT), Forward and Forward and

random primers, and reverse primers reverse primers
gene-specific primer for target gene and probe for
(GSP) enhancer (not included) target gene
solution (3 vials) (not included)

cDNA ready in cDNA ready in 35 min cDNA ready in 40 min cDNA ready in RT-qPCR data in RT-qPCR data
40 min for qPCR for qPCR for qPCR 40–90 min for qPCR 60–90 min in 60 min

© 2013 Bio-Rad Laboratories, Inc. Amplification Reagents and Plastics 9

 Reagents
Two-Step Reverse Transcription Reagents

iScript™ Advanced cDNA Synthesis Kit for RT-qPCR iScript Reverse Transcription Supermix
■■Increased qPCR data throughput and cost effectiveness from a single 20 µl for RT-qPCR
www.bio-rad.com/ reverse transcription (RT) reaction ■■ 1-tube format for simple and fast setup, and reduced pipetting variability
iscript
■■ Superior sensitivity and broad linear dynamic range for RT (7.5 µg–100 fg) ■■ Liquid format at –20°C offers superior stability and eliminates freeze/thaw cycle
■■  -tube kit (5x iScript reaction mix and iScript reverse transcriptase) for ease
2 ■■ Superior sensitivity and broad linear dynamic range for RT (1 µg–100 fg)
of use and reduced reaction setup time ■■  ptimized blend of oligo(dT) and random primers ensures complete and
O
■■  ptimized blend of oligo(dT) and random primers ensures complete and
O unbiased RNA sequence representation
unbiased RNA sequence representation ■■  Nase H+ MMLV reverse transcriptase (preblended with RNase inhibitor)
R
■■  Nase H+ MMLV reverse transcriptase (preblended with RNase inhibitor)
R delivers high sensitivity for real-time RT-qPCR and eliminates additional
delivers high sensitivity for real-time RT-qPCR and eliminates additional RNase H+ step
RNase H+ step ■■ Potent blend of RNaseA inhibitor protects RNA during setup and RT
■■ Potent blend of RNaseA inhibitor protects RNA during setup and RT ■■ Short 40 min protocol allows fast qPCR data generation
■■ Short 35 min protocol allows fast qPCR data generation For more information, download or request bulletin 6031.
For more information, download or request bulletin 6125.

35 104
— 1 µg RNA Average Cq SD CV, %
— 100 ng
103
104 100 ng 21.35 0.123 0.576
30 — 10 ng
RFU

— 1 ng 100 pg 31.56 0.147 0.465

102 — 100 pg
25 — 10 pg

RFU
RFU

— 1 pg
Cq

0 10 20 30 40
Cycles 103

20 103

102
15

–2 –1 0 1 2 3 4
log starting quantity
iScript advanced cDNA synthesis kit for RT-qPCR provides
superior sensitivity and a broad linear dynamic range for reverse
transcription. Total RNA (7.5 µg–1 pg) from HeLa cells was reverse
transcribed using the iScript advanced cDNA synthesis kit for RT-
qPCR in a 20 µl reaction. A tenfold dilution of generated cDNA was
used as template to amplify α-tubulin in a 10 µl qPCR reaction with
iQ™ SYBR® Green supermix on a CFX384™ real-time PCR detection
system. Standard curve R2 = 0.999, efficiency = 90.7%, slope = –3.57.
Cq, quantification cycle; RFU, relative fluorescence units.
0 10 20 30 40
Cycles
iScript reverse transcription supermix for RT-qPCR efficiently
reverse transcribes RNA over a broad linear dynamic range
for reliable gene expression analysis data. Different amounts of
HeLa cell RNA (amounts shown in inset) were reverse transcribed
and one-tenth of the resulting cDNA was used as a template to
amplify b-actin gene (~90 bp) in 20 µl qPCR reactions with iQ™
SYBR® Green supermix. Standard curve R 2 = 0.999, efficiency =
99.7%, slope = –3.33. RFU, relative fluorescence units.
10 20 30 40
Cycles
Excellent data reproducibility. PGK-1 mRNA (~160 bp), a gene that
encodes a glycolytic enzyme, was quantified using iScript reverse
transcription supermix for RT-qPCR both with 100 ng () and 100 pg ()
of input RNA. For each input RNA, 48 individual RT reactions were
performed and one-tenth of the resulting cDNA was used in the qPCR
reaction with SsoFast™ probes supermix. The gene expression analysis
data show excellent reproducibility both with high and low levels of input
target mRNA. The ~10 Cq difference for the 1,000-fold dilution of RNA
(100 ng–100 pg) demonstrates good reverse transcription efficiencies
across different input RNAs. Cq, quantification cycle; RFU, relative
fluorescence units.

Amplification Reagents and Plastics 10 Visit us on the Web at www.bio-rad.com.

 Reagents
Two-Step Reverse Transcription Reagents

iScript™ cDNA Synthesis Kit iScript Select cDNA Synthesis Kit

■■  -tube kit (5x iScript reaction mix and iScript reverse transcriptase) for
2 ■■  -tube kit (random primers, oligo[dT], 5x iScript Select reaction mix, iScript
5
ease of use and reduced reaction setup time reverse transcriptase, and gene-specific primer enhancer solution) www.bio-rad.com/
iscript
■■ Superior sensitivity and broad linear dynamic range for RT (1 µg–100 fg) ■■ Choice of priming strategy
■■  ptimized blend of oligo(dT) and random primers ensures complete and
O ■■ Reliable synthesis of long cDNA >6 kb in length
unbiased RNA sequence representation ■■ Superior sensitivity and broad linear dynamic range for RT (1 µg–1 pg)
■■  Nase H+ MMLV reverse transcriptase (preblended with RNase inhibitor)
R For more information, download or request bulletin 2894.
delivers high sensitivity for real-time RT-qPCR and eliminates additional
RNase H+ step

Reagents
■■ Potent blend of RNaseA inhibitor protects RNA during setup and RT
■■ Short 40 min protocol allows fast qPCR data generation
For more information, download or request bulletin 2894.

103 104
— 1 µg
— 100 ng 104
— 10 ng
— 1 ng
— 100 pg
— 10 pg

RFU
RFU

RFU

102 — 1 pg 103
103

10
0 5 10 15 20 25 30 35 40 45
Cycles
The iScript cDNA synthesis kit performs across a broad range
of concentrations. Input RNA (amounts shown in inset) was reverse
transcribed, and the resulting cDNA was amplified using iQ™ SYBR®
Green supermix. Standard curve r2 = 0.998, efficiency = 96.5%.
RFU, relative fluorescence units.
102
0 10 20 30 40
Cycles
iScript reagents provide potent RNaseA inhibition. iScript
reagents for RT-qPCR include an optimum blend of RNaseA
inhibitor for protecting RNA integrity. Reverse transcription was
performed using 0.1 pg of input RNA with iScript reagent alone (),
spiked with RNaseA (), or spiked with RNaseA without the
RNaseA inhibitor included in the reaction (). 18S rRNA (~70 bp)
was amplified using iQ™ SYBR® Green supermix. A significant Cq
delay was observed when the reaction included RNaseA but no
RNaseA inhibitor, which demonstrates potent RNaseA inhibition.
RFU, relative fluorescence units.
102
0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44
Cycles
iScript Select cDNA synthesis kit performs reliably over 6 orders
of magnitude using a gene-specific primer approach. Human
total RNA from 1 μg to 1 pg was reverse transcribed using the iScript
Select cDNA synthesis kit. One-tenth of the resulting cDNA was
used as a template to amplify b-actin gene with iQ™ SYBR® Green
supermix. Standard curve r = 1.000, efficiency = 92.2%. RFU, relative
fluorescence units.

© 2013 Bio-Rad Laboratories, Inc. Amplification Reagents and Plastics 11

 Reagents
One-Step RT-qPCR Reagents

iScript™ One-Step RT-PCR Kit with SYBR® Green Benefits of iScript one-step kits:
■■ For use on a broad range of real-time PCR instruments ■■  rovide powerful combination of iScript RNase H+ reverse transcriptase
P
www.bio-rad.com/ ■■ E xtremely sensitive detection (100 ng–1 pg) of input RNA and antibody-mediated hot-start iTaq™ DNA polymerase
iscript
A re ideal for rapid, high-throughput gene expression analysis
iScript One-Step RT-PCR Kit for Probes
■■

■■  or use with all types of hybridization probes, including dual-labeled

F ■■  erform cDNA synthesis and qPCR in 1 tube, minimizing handling
P
oligonucleotide probes, FRET probes, and molecular beacons and contamination risk
■■ E xtremely sensitive detection (1 µg–1 pg) of input RNA
For more information, download or request bulletin 3066.

PCR baseline-subtracted curve fit, RFU

PCR baseline-subtracted curve fit, RFU

104 103

103 102

102 10
0 2 4 6 8 10
12 14
16
18 20
22 24 26
28 30
32
34
36 38
40
42
44
46 48 0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46
Cycles Cycles
iScript™ one-step RT-PCR kit with SYBR® Green provides high reproducibility and iScript one-step RT-PCR kit for probes delivers unparalleled results over an extremely
sensitivity across a broad range of concentrations. Reactions were performed in triplicate, wide dynamic range. RNA (1 µg–100 fg) isolated from HeLa cells using the Aurum™ total
along with no-template controls, using GAPDH primers and 100 ng–100 fg total HeLa RNA. RNA kit was reverse transcribed and amplified using primers to b-actin and a FAM-labeled
Reactions were carried out on the iCycler iQ® real-time PCR detection system. Standard curve detection probe. Each dilution was performed in triplicate and RT-PCR was carried out on the
r = 1.000, efficiency = 95%, slope = –3.47. RFU, relative fluorescence units. iCycler iQ real-time PCR detection system. Standard curve r = 1.000, efficiency = 97.2%,
slope = –3.39. RFU, relative fluorescence units.

Ordering Information
Catalog # Description $
Two-Step Reverse Transcription Reagents One-Step RT-qPCR Reagents
170-8842 iScript Advanced cDNA Synthesis Kit for RT-qPCR, 50 x 20 μl reactions 170-8892 iScript One-Step RT-PCR Kit with SYBR Green, 50 x 50 μl reactions
170-8843 iScript Advanced cDNA Synthesis Kit for RT-qPCR, 250 x 20 μl reactions 170-8893 iScript One-Step RT-PCR Kit with SYBR Green, 200 x 50 μl reactions
170-8890 iScript cDNA Synthesis Kit, 25 x 20 μl reactions 170-8894 iScript One-Step RT-PCR Kit for Probes, 50 x 50 μl reactions
170-8891 iScript cDNA Synthesis Kit, 100 x 20 μl reactions 170-8895 iScript One-Step RT-PCR Kit for Probes, 200 x 50 μl reactions
170-8840 iScript Reverse Transcription Supermix for RT-qPCR, 25 x 20 μl reactions
170-8841 iScript Reverse Transcription Supermix for RT-qPCR, 100 x 20 μl reactions
170-8896 iScript Select cDNA Synthesis Kit, 25 x 20 μl reactions
170-8897 iScript Select cDNA Synthesis Kit, 100 x 20 μl reactions

Amplification Reagents and Plastics 12 Visit us on the Web at www.bio-rad.com.

Reagents —
Real-Time qPCR
■■  atented Sso7d fusion enzyme technology
P
delivers higher processivity and inhibitor
tolerance
■■  ntibody-mediated hot-start technology
A
enables instant polymerase activation and
superior specificity

Reagents
■■  hoice of fast, standard, or universal
C
cycling conditions
■■  ormulated for optimal performance on
F
a variety of real-time instruments
 Reagents ­
Real-Time qPCR Supermixes

Property SsoAdvanced™ iTaq™ Universal iQ™ Supermixes Application-Specific

Supermixes Supermixes Kits and Reagents
Tolerance for PCR inhibitors ••• – – –
Sensitive detection of low-level target genes ••• ••• •• •••
High efficiency even for difficult amplicons ••• •• •• •••
Broad range of reaction conditions ••• •• • •
Standard and fast cycling ••• ••• • •
Compatibility with any real-time instrument – ••• – –

Amplification Reagents and Plastics 14 Visit us on the Web at www.bio-rad.com.

 Reagents­
Real-Time qPCR Supermixes

SsoAdvanced™ Supermixes
■■ Superior performance even from compromised samples
■■ Tolerance for a broad range of reaction conditions and difficult amplicons www.bio-rad.com/
supermixes
■■ Optimal results from standard and fast PCR

iTaq™ Universal Supermixes

■■ Robust and sensitive qPCR data
■■ Instant polymerase activation
■■ Reliable results from standard and fast cycling conditions

Reagents
■■ Compatible with any real-time instrument

iQ™ Supermixes
■■ Reliable and reproducible qPCR performance
■■ qPCR with increased specificity
■■ Proven formulation for basic qPCR assays and needs
■■ Quick activation of antibody-mediated hot-start enzyme

Application-Specific Kits and Reagents

■■  uperior high resolution melt (HRM) data for single nucleotide
S
polymorphism (SNP) detection
■■ Accurate methylation detection
■■ Novel quantitative chromatin structure information using qPCR

© 2013 Bio-Rad Laboratories, Inc. Amplification Reagents and Plastics 15

 Reagents ­
SsoAdvanced™ Supermixes

SsoAdvanced™ SYBR® Green Supermix* Sso7d Polymerase

■■ Novel Sso7d fusion polymerase enables increased resistance to PCR

www.bio-rad.com/ inhibitors and higher processivity for dye-based real-time qPCR

supermixes
■■Robust formulation delivers maximum efficiency, sensitivity, and
reproducibility across a broad range of standard and fast cycling conditions
■■  ntibody-mediated hot-start technology and optimized buffer allow for instant
A
polymerase activation and rapid polymerization kinetics to enable fast PCR
■■  dvanced formulation tolerates a broad range of reaction conditions,
A
primer concentrations, and temperature ranges
For more information, download or request bulletin 6136.
* SsoAdvanced™ universal SYBR® Green supermix will be available soon and it will be The dsDNA binding protein, Sso7d, stabilizes the polymerase-template complex, increases
compatible with any real-time PCR instrument. processivity, and provides greater speed and reduced reaction times compared to
conventional DNA polymerases. Sso7d fusion polymerases are significantly more resistant to
PCR inhibitors, making the SsoAdvanced and SsoFast™ supermixes ideal choices for challenging
applications, such as direct qPCR, without the need for sample preparation.

105 104 105

38
700
36
600
34
–d(RFU)/dT

500
32

RFU
Cq

400
30 300
104 103 104
28 200
26 100
RFU

RFU
RFU

0
–12 –11 –10 –9 –8 65 70 75 80 85 90 95 18 19 20 21 22
log starting quantity Temperature, °C Cycles

103 102 103

102 10 102
0 10 20 30 40 0 10 20 30 40 0 10 20 30 40
Cycles Cycles Cycles

SsoAdvanced™ SYBR® Green supermix provides extreme
sensitivity in the detection of a single-copy target gene.
The cyclin gene was amplified and detected from fivefold serial
dilutions of 10 ng–80 pg (■) and 3.2 pg (■) human genomic DNA.
Standard curve R2 = 1 (for 10 ng–80 pg), cyclin efficiency = 103%.
Inset shows the standard curve for the various dilutions.
Cq, quantification cycle; RFU, relative fluorescence units.
SsoAdvanced™ SYBR® Green supermix demonstrates superior
inhibitor tolerance. The ADAR gene was amplified from HeLa cDNA in the
presence of water alone, or in the presence of a known PCR inhibitor,
Eagle’s minimum essential medium (EMEM) with fetal bovine serum (FBS; 0,
2.5, 5, 10, and 20%), added to SsoAdvanced™ SYBR® Green supermix (■)
or a traditional Taq DNA polymerase–based qPCR master mix (■).
SsoAdvanced™ SYBR® Green supermix showed quality amplification
in all reactions (EMEM with 20% FBS data shown) while the Taq DNA
polymerase–based qPCR master mix failed to amplify in all EMEM with FBS
combinations (shown in the inset melt curve). RFU, relative fluorescence units.
Exceptional reproducibility can be achieved with SsoAdvanced™
SYBR® Green supermix. Efficient discrimination and reliable
quantification can be obtained from a 1.33-fold serial dilution of input
template. The GAPDH gene was amplified from varying amounts of
HeLa cDNA (1 ng–136 pg). From left to right: 1 ng, 753 pg, 565 pg,
425 pg, 320 pg, 240 pg, 181 pg, and 136 pg. Standard curve R2 =
0.999, GAPDH efficiency = 96.2%. Inset is a magnified view showing
robust discrimination and reproducible amplification (six replicates for
each input amount). RFU, relative fluorescence units.

Amplification Reagents and Plastics 16 Visit us on the Web at www.bio-rad.com.

 Reagents­
SsoAdvanced™ Supermixes

SsoFast™ Probes Supermix Sso7d Polymerase

■■ Robust formulation allows simultaneous detection of up to 2 different

gene targets using fluorogenic probes with maximum efficiency, www.bio-rad.com/

sensitivity, and reproducibility across a broad range of standard and supermixes

fast cycling conditions

■■Novel Sso7d fusion polymerase enables increased resistance to PCR
inhibitors and higher processivity for probe-based real-time qPCR
■■  ntibody-mediated hot-start technology and optimized buffer allow for instant
A
polymerase activation and rapid polymerization kinetics to enable fast PCR

Reagents
For more information, download or request bulletin 5869.

The dsDNA binding protein, Sso7d, stabilizes the polymerase-template complex, increases
processivity, and provides greater speed and reduced reaction times compared to
conventional DNA polymerases. Sso7d fusion polymerases are significantly more resistant to
PCR inhibitors, making the SsoAdvanced and SsoFast supermixes ideal choices for challenging
applications, such as direct qPCR, without the need for sample preparation.

105 104
Plate # Start Time, PCR R2
hr Efficiency, %
1 0 97.0 0.999
Cq

104
103
2 14.0 97.6 0.999
RFU
RFU

log starting quantity

3 24.5 95.9 0.999
103 4 39.5 97.3 0.999
102
5 48.0 95.9 0.999

102
101
0 10 20 30 40 0 10 20 30 40
Cycles Cycles

SsoFast probes supermix delivers superior results for gene
expression analysis of two targets on the CFX96™ real-time
PCR detection system, with no difference in detection of a low-
expressing gene in duplex or simplex. cDNA from human liver
(100 ng) was used in each 20 µl reaction. (n) HEX-labeled GAPDH probe
duplex reaction; (n) Texas Red–labeled IL-2 probe duplex reaction;
(n) HEX-labeled GAPDH probe simplex reaction; (n) Texas Red–labeled
IL-2 probe simplex reaction. Total qPCR run time = 38 min. RFU, relative
fluorescence units.
Exceptional reproducibility can be achieved on the CFX384™
real-time PCR detection system with SsoFast probes supermix.
Efficient discrimination and reliable quantification can be obtained from
1.33-fold serial dilutions of input template. The GAPDH gene was
amplified from varying amounts of HeLa cDNA (1 ng–102 pg). From
left to right: () 1 ng, 565 pg, 320 pg, 181 pg, and 102 pg;
() 752 pg, 425 pg, 240 pg, and 136 pg. GAPDH efficiency =
91.5%, R2 = 0.997. Inset shows the standard curve for the various
dilutions. Total qPCR run time = 50 min. Cq, quantification cycle;
RFU, relative fluorescence units.
SsoFast probes supermix maintains exceptional stability on
the high-throughput CFX automation system. Tenfold serial
dilutions of 100 ng–1 pg cDNA from human spleen were used in
each 20 µl reaction to detect GAPDH. All reactions were assembled
and loaded onto the CFX automation system. The following cycling
conditions were used: 95°C for 10 min, followed by 35 cycles
of 95°C for 15 sec and 60°C for 60 sec. After each plate run,
an additional hold time was introduced to prolong the total time
between plates. Results for five plates (0–48 hr) are shown.

© 2013 Bio-Rad Laboratories, Inc. Amplification Reagents and Plastics 17

 Reagents ­
iTaq™ Universal Supermixes

iTaq™ Universal SYBR® Green Supermix

■■  dvanced 2x ready-to-use supermix, formulated to deliver robust qPCR
A
www.bio-rad.com/ results with superior sensitivity, efficiency, and specificity
supermixes
■■  ptimized buffer allows consistent results using both standard and fast
O
cycling protocols
■■  ntibody-mediated iTaq DNA polymerase enables fast activation and
A
superior specificity in qPCR
■■ Formulation developed for optimal results on any real-time PCR instrument

105

104 104
RFU

103 104
RFU

RFU
RFU

17 18 19 20 21 22 23 24 25 26 103
Cycles
103 103

102

102 102 10
0 5 10 15 20 25 30 35 0 10 20 30 40 0 10 20 30 40
Cycles Cycles Cycles

Exceptional reproducibility can be achieved with iTaq™
universal SYBR® Green supermix. Efficient discrimination and
reliable quantification can be obtained from a 1.33-fold serial
dilution of input template. The human b-actin gene was amplified
from varying amounts of HeLa cDNA (1 ng–136 pg). From left to
right: 1 ng, 753 pg, 565 pg, 425 pg, 320 pg, 240 pg, 181 pg, and
136 pg. Inset is a magnified view showing robust discrimination
and reproducible amplification (six replicates for each input
amount). RFU, relative fluorescence units.
iTaq™ universal SYBR® Green supermix provides reliable gene
expression data. The human b-actin gene was amplified from HeLa cDNA
(100 ng–100 fg) using a CFX96™ real-time PCR detection system. iTaq™
universal SYBR® Green supermix produced greater than 90% efficiency
over 6 orders of linear dynamic range. Standard curve R2 = 0.999, ß-actin
efficiency = 92.6%, slope = –3.51. RFU, relative fluorescence units.
iTaq™ universal SYBR® Green supermix allows robust
amplification of genomic DNA. The human GAPDH gene was
amplified from human genomic DNA (50 ng–5 pg) using a CFX96
real-time PCR detection system. iTaq™ universal SYBR® Green
supermix produced a GAPDH efficiency of 108.1% over several orders
of linear dynamic range. Standard curve R2 = 0.993, slope = –3.14.
RFU, relative fluorescence units.

Amplification Reagents and Plastics 18 Visit us on the Web at www.bio-rad.com.

 Reagents­
iTaq™ Universal Supermixes

iTaq Universal Probes Supermix

■■  ptimized buffer allows consistent results for simplex and duplex reactions
O
using both standard and fast cycling protocols www.bio-rad.com/
supermixes
■■Advanced 2x ready-to-use supermix, formulated to deliver robust
qPCR results with superior sensitivity, efficiency, and specificity
■■  ntibody-mediated iTaq DNA polymerase enables fast activation and
A
superior specificity in qPCR
■■ Formulation developed for optimal results on any real-time PCR instrument

Reagents
104 104 RFU 104

102
103 103
RFU

RFU

RFU
18 20 22 24 26 103
Cycles

102 102

102
0 10 20 30 40 0 10 20 30 40 0 10 20 30 40
Cycles Cycles Cycles

iTaq universal probes supermix is excellent for gene expression
analysis. The human b-actin gene was amplified from HeLa cDNA
(100 ng–100 fg) using FAM-labeled probes on a CFX96™ real-time PCR
detection system. iTaq universal probes supermix produced a b-actin
efficiency of 94.3% over 6 orders of linear dynamic range. Standard
curve R2 = 0.999, slope = –3.47. RFU, relative fluorescence units.
Exceptional reproducibility can be achieved with iTaq universal
probes supermix. Efficient discrimination and reliable quantification
can be obtained from a 1.33-fold serial dilution of input template. The
human b-actin gene was amplified from varying amounts of HeLa
cDNA (1 ng–136 pg). From left to right: 1 ng, 753 pg, 565 pg, 425 pg,
320 pg, 240 pg, 181 pg, and 136 pg. Inset is a magnified view showing
robust discrimination and reproducible amplification (six replicates for
each input amount). RFU, relative fluorescence units.
iTaq universal probes supermix allows accurate detection of
low-abundance targets. The IL-1b gene was amplified from HeLa
cDNA (100, 10, 1, and 0.1 ng) using FAM-labeled probes on a CFX96
real-time PCR detection system. iTaq universal probes supermix
showed sensitive detection of the IL-1b gene even with very low
input cDNA. RFU, relative fluorescence units.

© 2013 Bio-Rad Laboratories, Inc. Amplification Reagents and Plastics 19

 Reagents­­
iQ™ Supermixes

iQ™ SYBR® Green Supermix

■■  nalysis of low-, medium-, and high-abundance target genes with superior
A
www.bio-rad.com/ sensitivity and efficiency
supermixes
■■  ormulated for maximum SYBR® Green I stability and performance in
F
a wide variety of real-time PCR instruments
■■  ntibody-mediated hot-start polymerase for quick activation and
A
increased specificity
For more information, download or request bulletin 2764.

104

PCR baseline-subtracted curve fit, RFU

PCR baseline-subtracted curve fit, RFU

103

103
102

0 5 10 15 20 25 30 35 40
Cycles
iQ™ SYBR® Green supermix generates precise, quantitative results.
A fivefold dilution series (50 ng–80 pg) of human genomic DNA was amplified
using the supermix, primers, and a probe specific to the IL-1b gene. Triplicate
reactions at each concentration were amplified along with no-template
controls on the iCycler iQ® real-time PCR detection system. Standard curve
r = 0.999, efficiency = 97.6%, slope = –3.38. RFU, relative fluorescence units.
0 5 10 15 20 25 30 35 40 44
Cycles
iQ™ SYBR® Green supermix and the iScript™ cDNA synthesis kit show
consistently high specificity over a broad dynamic range of cDNA.
Serial dilutions (1 µg–1 pg) of HeLa total RNA were reverse transcribed,
and the resulting cDNA was amplified using primers specific to the b-actin
gene. Triplicate reactions at each concentration were amplified along with
no-template controls on the iCycler iQ real-time PCR detection system. The
consistent spacing of the curves reflects accurate reverse transcription and
amplification. Standard curve r = 0.997, efficiency = 89.1%, slope = –3.62.
RFU, relative fluorescence units.

Amplification Reagents and Plastics 20 Visit us on the Web at www.bio-rad.com.

 Reagents­
iQ™ Supermixes

iQ Supermix iQ Multiplex Powermix

■■ Maximum efficiency and sensitivity for qPCR using fluorogenic probes ■■ Robust supermix formulated for sensitive and efficient multiplex qPCR
■■  eliable amplification over a wide dynamic range of human genomic and
R ■■  eliable quantification of up to 4 targets (expression levels can vary up
R www.bio-rad.com/
supermixes
plasmid DNA concentrations to 106-fold between target genes) or up to 5 targets
■■ Contains antibody-mediated hot-start iTaq™ DNA polymerase for quick ■■  inearity over 6 orders of magnitude of input cDNA and 4 orders of
L
activation and increased specificity magnitude of input genomic DNA
For more information, download or request bulletin 2764. ■■  uitable for a wide variety of applications, including gene expression
S
analysis, SNP genotyping, SNP analysis, GMO detection, and viral
load detection

Reagents
For more information, download or request bulletin 5348.

103

103
RFU

102 RFU

102

0 5 10 15 20 25 30 35 40 45 50 0 5 10 15 20 25 30 35 40 45
Cycles Cycles

iQ supermix provides sensitive real-time detection over 8 orders
of magnitude. Tenfold dilutions of a plasmid containing 109–10 copies
of the a-tubulin gene were amplified using iQ supermix and a FAM-
labeled hybridization probe for detection. Eight replicates at each
concentration were amplified along with no-template controls on the
MyiQ™ real-time PCR detection system. Standard curve r = 0.999,
efficiency = 98.2%, slope = –3.36. RFU, relative fluorescence units.
iQ multiplex powermix produces highly reliable qPCR results
for up to five targets in a single tube, with no difference in
detection of a low-expressing gene in multiplex or singleplex.
One-tenth of a 1 µg cDNA synthesis reaction of human thymus total
RNA was used in each 20 µl reaction. FAM-labeled b-actin probe (),
Cy5-labeled a-tubulin probe (), HEX-labeled GAPDH probe (),
TAMRA-labeled cyclophilin probe (), Texas Red–labeled IL-2 probe ().
RFU, relative fluorescence units.

© 2013 Bio-Rad Laboratories, Inc. Amplification Reagents and Plastics 21

 Reagents­­
Application-Specific Kits and Reagents

Precision Melt Supermix

■■ 
Optimized formulation containing EvaGreen dye delivers robust PCR and
www.bio-rad.com/ high resolution melt (HRM) performance
supermixes
■■  ensitive and effective discrimination of all 4 SNP classes across a broad
S
range of amplicons
■■ Accurate detection of CpG methylation status for epigenetic studies
■■ Exceptional room temperature stability for high-throughput HRM studies
■■ Reliable performance on any HRM-capable thermal cycler
For more information, download or request bulletin 6137.

A B

0.20 0.02 0.0

–0.1
0.15 0.00
Difference RFU

–0.2
Difference RFU

Difference RFU
0.10 –0.02
–0.3
–0.04
0.05 –0.4

–0.06 –0.5
0.00

–0.08 –0.6
–0.05
78 79 80 81 82 83 75 76 77 78 79 80 74 76 78 80 82 84
Temperature, °C Temperature, °C Temperature, °C

Precision melt supermix delivers robust HRM for SNPs. Discrimination of class I and IV SNP genotypes are shown in panels A Accurate methylation detection with precision melt supermix. Mixtures
and B, respectively. Class I (A to G substitution) and class IV (A to T substitution) SNP genotypes from mouse genomic DNA were analyzed of methylated and unmethylated human genomic DNA of varying ratios were
using precision melt supermix. Wild type (■), heterozygote (■), and homozygous mutant (■) are shown in the difference plots normalized analyzed using HRM on a CFX384 real-time PCR detection system. Increasing
to wild-type samples. HRM analysis was performed on a CFX384™ real-time PCR detection system and genotypes were automatically amounts of methylated DNA (■, 0%; ■, 2%; ■, 5%; ■, 50%; ■, 75%; ■, 95%;
assigned by Precision Melt Analysis™ software. Amplification was carried out for 35 cycles. Total run time including melt curve = 150 min. and ■, 100%) were analyzed for methylation of the human RARB2 gene. The
RFU, relative fluorescence units. genomic region contains 7 CpG sites and is 88 base pairs in length. Total run
time including melt curve = 190 min. RFU, relative fluorescence units.

Amplification Reagents and Plastics 22 Visit us on the Web at www.bio-rad.com.

 Reagents­
Application-Specific Kits and Reagents

EpiQ™ Chromatin Analysis Kit EpiQ™ Chromatin SYBR® Green Supermix

■■  ovel technique generates quantitative chromatin structure information
N ■■  obust formulation delivers superior sensitivity and efficiency for
R
with strong correlation to gene expression levels qPCR from genomic DNA templates www.bio-rad.com/
epiq
■■  uantitative assessment of chromatin structure of target genes in
Q ■■  rotocol is optimized for difficult real-time qPCR reactions for high GC
P
cultured cells amplicons
■■  it discriminates open, actively transcribed chromatin regions from closed,
K ■■  upermix contains fluorescein and ROX and is compatible with all
S
transcriptionally silent regions real-time PCR instruments except Applied Biosystems 7000, 7300,
■■ In situ chromatin digestion, genomic DNA purification, and real-time PCR 7700, and 7900 models (additional ROX can be added by ordering
all combined in one workflow the dye separately, catalog #172-5858)

Reagents
■■  hort assay time — assessment of chromatin structure can be
S For more information, download or request bulletin 6020.

accomplished in less than 6 hr

■■  mall sample requirement — as little as 5 x 104 cells are required to
S
perform analysis
For more information, download or request bulletin 6020.

A. HBB — Reference Gene (epigenetically silenced) B. GAPDH — Target Gene (constitutively expressed)
104 — No nuclease 104 — No nuclease
— With nuclease — With nuclease

Closed chromatin
103 103
RFU

RFU
Open chromatin
102 102
∆Cq ref = 0.58 ∆Cq target = 8.08

101 101

Chromatin consists of DNA spooled around complexes of histone
protein molecules called nucleosomes ().
0 10 20 30 40 0 10 20 30 40
Cycles Cycles
The EpiQ chromatin analysis kit utilizes nuclease accessibility to discriminate open vs. closed chromatin regions.
Amplification of proximal promoter regions for the epigenetically silenced HBB (reference) gene or the constitutively expressed
GAPDH (target) gene was carried out in HeLa cells using the EpiQ kit and EpiQ™ chromatin SYBR® Green supermix on the
CFX96™ real-time PCR detection system. A, closed chromatin regions were protected from nuclease digestion and remained
intact prior to amplification, resulting in minimal quantification cycle (Cq) delays (ΔCq = 0.58) following nuclease treatment; B, open
chromatin regions were susceptible to nuclease digestion and were unavailable for amplification, leading to significant Cq delays
(ΔCq = 8.08) after nuclease treatment. A comparison of ΔCqs with the amplification efficiencies for each gene target factored in
was used to determine the accessibility of the target gene, calculated to be >99% for GAPDH. RFU, relative fluorescence units.

© 2013 Bio-Rad Laboratories, Inc. Amplification Reagents and Plastics 23

 Reagents­
Standard PCR Reagents

iTaq™ DNA Polymerase iProof™ High-Fidelity DNA Polymerase

■■ 
Antibody-mediated hot-start DNA polymerase for quick 3 min activation ■■  high-fidelity DNA polymerase with 52-fold more accuracy than Taq
A
www.bio-rad.com/ at 95°C DNA polymerase
pcrreagents
■■  olymerase prevents nonspecific amplification and primer-dimers in both
P ■■ Unique Pyrococcus-like proofreading enzyme is fused to a dsDNA binding
PCR and real-time PCR applications protein, Sso7d
For more information, download or request bulletin 2779. ■■  ong and fast PCR applications — fragments up to 37 kb are amplified
L
in less time (15–30 sec/kb) and with less enzyme (0.25–1 U/reaction)
dNTP Mix
■■ Formulated for consistency and higher efficiency in PCR and real-time PCR
■■  onvenient 2x supermix is available for iProof polymerase and buffer
C
for GC-rich templates
■■  obust dNTP solution withstands multiple rounds of freeze-thawing and
R
For more information, download or request bulletin 5211.
temperature cycling

BAC DNA (37 kb) Human genomic DNA (28 kb)

35 min iProof high-fidelity DNA polymerase
1 kb 2 hr
Common high-fidelity polymerase – 48 kb
2 hr
Taq polymerase 15 22 28
Template length

32 37 – 23 kb
1.5 hr 10 20 22 25 7.9
8 kb 9.5 hr
5.5 hr 5

2 hr 20 min
15 kb 16.5 hr
(Failed amplification)
Total protocol time
iProof high-fidelity DNA polymerase demonstrates unrivaled speed, leading to dramatically
shorter overall reaction times. The reaction protocol for iProof polymerase was compared to the
recommended protocols for two competing polymerases. Each protocol was designed to amplify 1,
8, and 15 kb products in 30 cycles. Reactions with iProof polymerase used a two-step protocol with a
combined annealing and extension step, while the other reactions used three-step protocols with the
minimum recommended extension times. Overall reaction times include temperature ramping times.
3.8
2.9
iProof high-fidelity DNA polymerase amplifies long templates with high yields. Left, various
fragments up to 37 kb in length were amplified from BAC DNA using a combined annealing/
extension step of 10 min per cycle and 30 U/ml of iProof polymerase. Right, various sequences up
to 28.8 kb were amplified directly from human genomic DNA using 30 U/ml of iProof polymerase in
GC buffer with a combined annealing/extension time of 10 min per cycle.

Amplification Reagents and Plastics 24 Visit us on the Web at www.bio-rad.com.

 Reagents­
Real-Time qPCR Reagents Selection Guide

SYBR® Green / EvaGreen Supermixes Probes Supermixes One-Step Kits for RT-
qPCR
SsoAdvanced™ iTaq™ iQ™ SYBR® SsoFast™ EpiQ™ SsoFast iTaq iQ iQ iScript™ iScript
SYBR® Universal Green EvaGreen® Chromatin Probes Universal Supermix Multiplex One-Step One-Step
Green SYBR® Green Supermix Supermix SYBR® Green Supermix Probes Powermix RT-PCR Kit RT-PCR Kit
Supermix* Supermix Supermix Supermix with SYBR® for Probes
Real-Time PCR Instrument Green
Bio-Rad
CFX96™, CFX96 Touch™, CFX96
Touch Deep Well™, CFX384™, ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔
CFX384 Touch™, CFX Connect™
iQ™, iQ™5, MyiQ™, MyiQ™2 ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔
MiniOpticon™, DNA Engine
✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔

Reagents
Opticon® I and II
Applied Biosystems
StepOne/StepOne Plus ◆ ✔ ◆ ◆ ✔ ◆ ✔ ◆ ◆ ◆ ◆
7500, ViiA 7 ✔ ✔ ✔
7000, 7300, 7700, 7900HT ✔ ✔
Stratagene

Mx3000P, 3005P, 4000 ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔

Eppendorf
Mastercycler ep
realplex 2 or 4
✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔
QIAGEN/Corbett
Rotor-Gene 3000, 6000, Q ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔
Roche

LightCycler 480 ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔
LightCycler 1.0, 1.5, 2.0 ▲ ▲ ▲ ▲ ✔ ▲ ▲ ▲ ▲ ▲ ▲
Idaho Technology
LightScanner HR-1 ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔
LightScanner 32 ▲ ▲ ▲ ▲ ✔ ▲ ▲ ▲ ▲ ▲ ▲

✔ Recommended for use as is ◆ ROX reference setting must be turned “off” ▲ BSA must be added according to instrument specifications
* SsoAdvanced™ universal SYBR® Green supermix will be available soon.

© 2013 Bio-Rad Laboratories, Inc. Amplification Reagents and Plastics 25

 Reagents ­
Ordering Information

Ordering Information
Catalog # Description $
SsoAdvanced Supermixes
172-5260 SsoAdvanced SYBR Green Supermix, 2 ml (2 x 1 ml vials), 200 x 20 μl reactions
172-5261 SsoAdvanced SYBR Green Supermix, 5 ml (5 x 1 ml vials), 500 x 20 μl reactions
172-5262 SsoAdvanced SYBR Green Supermix, 10 ml (10 x 1 ml vials), 1,000 x 20 μl reactions
172-5264 SsoAdvanced SYBR Green Supermix, 25 ml (5 x 5 ml vials), 2,500 x 20 μl reactions
172-5265 SsoAdvanced SYBR Green Supermix, 50 ml (10 x 5 ml vials), 5,000 x 20 μl reactions
172-5230 SsoFast Probes Supermix, 2 ml (2 x 1 ml vials), 200 x 20 μl reactions
172-5231 SsoFast Probes Supermix, 5 ml (5 x 1 ml vials), 500 x 20 μl reactions
172-5232 SsoFast Probes Supermix, 10 ml (10 x 1 ml vials), 1,000 x 20 μl reactions
172-5233 SsoFast Probes Supermix, 20 ml (20 ml bottle), 2,000 x 20 μl reactions
iTaq Universal Supermixes
172-5120 iTaq Universal SYBR Green Supermix, 2 ml (2 x 1 ml vials), 200 x 20 μl reactions
172-5121 iTaq Universal SYBR Green Supermix, 5 ml (5 x 1 ml vials), 500 x 20 μl reactions
172-5122 iTaq Universal SYBR Green Supermix, 10 ml (10 x 1 ml vials), 1,000 x 20 μl reactions
172-5124 iTaq Universal SYBR Green Supermix, 25 ml (5 x 5 ml vials), 2,500 x 20 μl reactions
172-5125 iTaq Universal SYBR Green Supermix, 50 ml (10 x 5 ml vials), 5,000 x 20 μl reactions
172-5130 iTaq Universal Probes Supermix, 2 ml (2 x 1 ml vials), 200 x 20 μl reactions
172-5131 iTaq Universal Probes Supermix, 5 ml (5 x 1 ml vials), 500 x 20 μl reactions
172-5132 iTaq Universal Probes Supermix, 10 ml (10 x 1 ml vials), 1,000 x 20 μl reactions
172-5134 iTaq Universal Probes Supermix, 25 ml (5 x 5 ml vials), 2,500 x 20 μl reactions
172-5135 iTaq Universal Probes Supermix, 50 ml (10 x 5 ml vials), 5,000 x 20 μl reactions
iQ Supermixes
170-8880 iQ SYBR Green Supermix, 2.5 ml (2 x 1.25 ml vials), 100 x 50 μl reactions
170-8882 iQ SYBR Green Supermix, 12.5 ml (10 x 1.25 ml vials), 500 x 50 μl reactions
170-8884 iQ SYBR Green Supermix, 25 ml (20 x 1.25 ml vials), 1,000 x 50 μl reactions
170-8885 iQ SYBR Green Supermix, 50 ml (50 ml bottle), 2,000 x 50 μl reactions
170-8886 iQ SYBR Green Supermix, 25 ml (5 x 5 ml vials), 1,000 x 50 μl reactions
170-8887 iQ SYBR Green Supermix, 50 ml (10 x 5 ml vials), 2,000 x 50 μl reactions
170-8860 iQ Supermix, 2.5 ml (2 x 1.25 ml vials), 100 x 50 μl reactions
170-8862 iQ Supermix, 12.5 ml (10 x 1.25 ml vials), 500 x 50 μl reactions
170-8864 iQ Supermix, 25 ml (20 x 1.25 ml vials), 1,000 x 50 μl reactions
172-5848 iQ Multiplex Powermix, 1.25 ml (1 x 1.25 ml vial), 50 x 50 μl reactions
172-5849 iQ Multiplex Powermix, 5 ml (4 x 1.25 ml vials), 200 x 50 μl reactions

Amplification Reagents and Plastics 26 Visit us on the Web at www.bio-rad.com.

 Reagents ­
Ordering Information

Ordering Information
Catalog # Description $
Application-Specific Kits and Reagents
172-5110 Precision Melt Supermix, 2 ml (2 x 1 ml vials), 200 x 20 μl reactions
172-5112 Precision Melt Supermix, 10 ml (10 x 1 ml vials), 1,000 x 20 μl reactions
172-5400 EpiQ Chromatin Analysis Kit, 50 preparations
172-5401 EpiQ Chromatin Analysis Kit, 100 preparations
172-5404 EpiQ Chromatin SYBR Green Supermix, 5 ml (5 x 1 ml vials), 500 x 20 μl reactions
172-5405 EpiQ Chromatin SYBR Green Supermix, 10 ml (10 x 1 ml vials), 1,000 x 20 μl reactions
Standard PCR Reagents
170-8870 iTaq DNA Polymerase, 250 U, 5 U/μl
170-8875 iTaq DNA Polymerase, 5,000 U, 5 U/μl
170-8874 dNTP Mix, 200 μl

Reagents
172-5300 iProof High-Fidelity DNA Polymerase, 20 U, 2 U/μl
172-5301 iProof High-Fidelity DNA Polymerase, 100 U, 2 U/μl
172-5302 iProof High-Fidelity DNA Polymerase, 500 U, 2 U/μl
172-5310 iProof HF Master Mix, 0.04 U/μl, 100 x 50 μl reactions
172-5320 iProof GC Master Mix, 0.04 U/μl, 100 x 50 μl reactions
172-5858 ROX Passive Reference Dye, 0.5 ml
Additional Real-Time qPCR Supermixes*
172-5200 SsoFast EvaGreen Supermix, 2 ml (2 x 1 ml vials), 200 x 20 μl reactions
172-5201 SsoFast EvaGreen Supermix, 5 ml (5 x 1 ml vials), 500 x 20 μl reactions
172-5202 SsoFast EvaGreen Supermix, 10 ml (10 x 1 ml vials), 1,000 x 20 μl reactions
172-5203 SsoFast EvaGreen Supermix, 20 ml (20 ml bottle), 2,000 x 20 μl reactions
172-5204 SsoFast EvaGreen Supermix, 25 ml (5 x 5 ml vials), 2,500 x 20 μl reactions
172-5205 SsoFast EvaGreen Supermix, 50 ml (10 x 5 ml vials), 5,000 x 20 μl reactions

* There are additional supermixes available. For more information, go to www.biorad.com/supermixes.

© 2013 Bio-Rad Laboratories, Inc. Amplification Reagents and Plastics 27

PrimePCR™
Assays and Panels
■■  redesigned assays — approximately 20,000
P
human primer assays for SYBR® Green gene
expression analysis are available packaged
in vials of 200, 1,000, or 2,500 reactions.
Genome-wide mouse assays are coming soon
■■  redesigned panels — a large selection of
P
predesigned disease- and pathway-specific
panels is available
■■  xperimental controls — control assays
E
are available for reverse transcription,
RNA quality, genomic DNA contamination,
and PCR performance
■■  NA templates — synthetic DNA templates are
D
designed to give a positive real-time PCR result
when used with the corresponding gene assay
■■  ustom assays — user-defined primer
C
sequences can be ordered in vials of 200,
1,000, or 2,500 reactions
■■  ustom plates — custom-configured
C
96- and 384-well PCR plates can be ordered
 PrimePCR™
Assay Design and Validation

Criteria Used when Designing the We have collaborated with Biogazelle, leaders in real-time PCR, to design and
experimentally validate SYBR® Green–based gene expression assays
SYBR® Green–Based qPCR Assays
for human protein-coding genes.
■■ High assay specificity without use of a probe
■■  ssays provide confidence in results while eliminating time-consuming
A
■■ Avoided SNPs in target regions design and optimization steps
■■ Designed intron-spanning assays whenever possible ■■  IQE compliance (Bustin et al. 2009) made easy — validation information
M
■■ Avoided secondary structures in primer annealing sites for each assay is available at www.bio-rad.com/PrimePCR
■■ Maximized fraction of transcript isoforms being detected
■■ Compatibility with standard assay conditions
■■ Used latest release of genome builds and annotation databases
Assay Performance Standards
Sensitivity Accurate detection of 20 copies
Specificity 
Amplicon sequence validated with next-
generation sequencing; minimal primer-dimer
formation and genomic DNA cross-reactivity
Amplification efficiency 90–110%
Linear dynamic range Minimum of 6 orders of magnitude; detection
of a synthetic template standard curve from

PrimePCR
20 to 20 million copies
R2 >0.98

For more information, download bulletin 6262.

Assays were validated using iScript™
advanced cDNA synthesis kit for
RT-qPCR and SsoAdvanced™
SYBR® Green supermix on
an automated CFX384 Touch™
real-time PCR detection system.

For ordering information, please visit

www.bio-rad.com/PrimePCR.

© 2013 Bio-Rad Laboratories, Inc. Amplification Reagents and Plastics 29

 PrimePCR™
Pathway and Collection Panels

Pathway-Focused Analysis
Bio-Rad Laboratories has partnered with GeneGo, a Thomson
Reuters business and leading provider of systems biology tools,
to offer a complete pathway solution for real-time PCR–based
gene expression analysis. As part of the collaboration,
Thomson Reuters provided interactive pathway maps for
the canonical pathways.
These pathways were used to design panels of real-time PCR
assays specifically tailored to the top-ranked gene targets for
differential gene expression analysis.

Target Ranking — The Most Biologically

Relevant Assays
Gene assays present on PrimePCR pathway panels have been
prioritized based on three main criteria:
■■  ow often a gene changes expression level in
H
transcriptome studies
■■  ow much attention was paid to this gene in the overall
H
scientific research
■■  ow interesting the scientific community found this
H
gene in the last 2 years

Broad Target Exploration

PrimePCR collection panels represent the top-ranked gene targets
for differential gene expression analysis across a set of related
canonical pathways, allowing for a more general survey of gene
targets across a biological process or group. Panels were designed
for each of the general biological categories.

For more information, download bulletin 6263.

Note: Number in parenthesis refers to the number of pathways in a given category.

For ordering information, please visit www.bio-rad.com/
PrimePCR_Pathways.

Amplification Reagents and Plastics 30 Visit us on the Web at www.bio-rad.com.

 PrimePCR™
Custom Plates

Design a Custom Plate with PrimePCR Assays

■■  6- or 384-well plates are available for every major
9
real-time PCR instrument
■■  ustomize your plate design layout or use a suggested
C
plate template
■■ Include PrimePCR assays and/or custom assays on
your custom-designed plate

PrimePCR
For ordering information, please visit www.bio-rad.com/
PrimePCR_Custom_Plates.

© 2013 Bio-Rad Laboratories, Inc. Amplification Reagents and Plastics 31

PCR Plastic Consumables
■■  recisely manufactured for optimal fit and
P
cycling performance
■■  roduced in Class 10,000 or 100,000 cleanroom
P
environment
■■  ertified to be free of DNase, RNase, and human
C
genomic DNA
■■  xtremely uniform wells reduce well-to-well
E
variability in real-time PCR
■■ Warp-free Hard-Shell® plates are designed for
optimum performance with automation
 PCR Plastic Consumables
Instrument Compatibility

Tubes
Individual High-Profile Strips High-Profile Strips Low-Profile
Catalog# TBI-0201, TFI-0201, TWI-0201 TBS-xxxx, TBC-xxxx TLS-xxxx
Thermal Cycler

Bio-Rad® C1000™, C1000 Touch™, S1000™ ✔ ▲ ✔

Bio-Rad® DNA Engine®,
DNA Engine Tetrad®,
DNA Engine Tetrad 2, ✔ ▲ ✔
DNA Engine Dyad®,
Dyad Disciple™, PTC-100®
Bio-Rad® T100™, MyCycler™ ✔ ✔
Bio-Rad® iCycler ® ✔ ✔
Bio-Rad® MJ Mini™ ✔ ▲ ✔
Applied Biosystems 0.2 ml tube cyclers
✔ ✔
(2720, 9700, Veriti)

Applied Biosystems 0.1 ml tube cyclers

✔
(9800 fast, Veriti fast)
Eppendorf Mastercycler series ✔ ▲ ✔
Real-Time PCR Instrument
Bio-Rad® CFX96™, CFX96 Touch™, CFX96
✔
Touch Deep Well™, CFX Connect™
Bio-Rad® iCycler iQ®, iQ™5, MyiQ™, MyiQ™2 ✔
Bio-Rad® Chromo4™ ▲ ✔
®
Bio-Rad DNA Engine Opticon®, Opticon 2 ✔
Bio-Rad® MiniOpticon™* ✔
Applied Biosystems standard systems
✔
(7300, 7500, 7900HT)
Applied Biosystems fast systems (7500 fast,
✔
7900HT fast, StepOne, StepOnePlus)
Eppendorf Mastercycler ep realplex ▲ ✔
Stratagene (Agilent) Mx series ✔
QIAGEN/Corbett Rotor-Gene ✔

Plastics
✔ Recommended ▲ Compatible
* The MiniOpticon real-time PCR detection system is factory calibrated for white tubes and white-well plates. White plastics are recommended due to their superior signal-to-noise ratio. Using clear tubes or clear-well plates on
this instrument will require user calibration.

© 2013 Bio-Rad Laboratories, Inc. Amplification Reagents and Plastics 33

 PCR Plastic Consumables
Instrument Compatibility

Instrument Compatibility of PCR Plastic Consumables (cont.)

96- and 48-Well Plates 384-Well Plates

Hard-Shell ® Hard-Shell Hard-Shell Hard-Shell Multiplate™ Multiplate iQ™ Hard-Shell Hard-Shell

Semi-Skirted Skirted Semi-Skirted 480 Unskirted Unskirted Semi-Skirted Standard 480
High-Profile Low-Profile Low-Profile High-Profile Low-Profile High-Profile

Catalog # HSS-9xxx HSP-9xxx HSL-9xxx HSR-99xx MLP-xxxx MLL-xxxx 223-9441 HSP-3xxx HSR-48xx

Thermal Cycler

Bio-Rad® C1000™, C1000 Touch™, S1000™ ▲ ✔ ▲ ▲ ▲ ▲ ✔ ▲

Bio-Rad® T100™ ✔ ▲ ▲

Bio-Rad® DNA Engine®, DNA Engine Tetrad®,

DNA Engine Tetrad 2, DNA Engine Dyad®, ▲ ✔ ▲ ▲ ▲ ✔ ▲
Dyad Disciple™, PTC-100 ®

Bio-Rad® iCycler ®, MyCycler™ ✔ ✔ ▲

Except MyCycler

®
Bio-Rad MJ Mini ™
▲ ✔

Applied Biosystems 0.2 ml tube cyclers

✔ ▲ ▲
(2720, 9700, Veriti)
Applied Biosystems 0.1 ml tube cyclers
✔ ✔
(9800 fast, Veriti fast)
Applied Biosystems 384-well cyclers
✔ ▲
(9700, Veriti)
Eppendorf Mastercycler series ▲ ✔ ▲ ▲ ▲ ▲ ✔ ▲

Real-Time PCR Instrument

Bio-Rad® CFX96™, CFX96 Touch™,

✔ ▲ ▲ ✔ ▲
CFX Connect™, CFX384™,* CFX384 Touch™ *

CFX96 Touch Deep Well™ ▲ ✔

Bio-Rad® iCycler iQ®, iQ™5, MyiQ™, MyiQ™2 ▲ ▲ ✔

Bio-Rad® Chromo4™ ▲ ✔ ▲ ▲ ▲

Bio-Rad® DNA Engine Opticon®,

✔ ▲
Opticon 2, MiniOpticon™* Except MiniOpticon

Applied Biosystems standard systems

✔ ▲ ▲ ✔ ▲
(7300, 7500, 7900HT, ViiA 7)
Except ViiA 7 Except 7900HT Except 7900HT

Applied Biosystems fast systems

✔ ▲ ✔ ▲
(7500 fast, 7900HT fast, StepOne, StepOnePlus, ViiA 7 fast) Except StepOne, StepOnePlus, 7900 fast Except 7900 fast

Eppendorf Mastercycler ep realplex ▲ ✔ ▲ ▲ ▲ ▲

Stratagene (Agilent) Mx series ✔ ▲ ▲

Roche LightCycler 480 ✔ ✔

Other Instruments

Applied Biosystems DNA sequencers

✔ ▲ ✔
(3100, 3700, 3730)

Idaho Technology LightScanner ✔ ▲ ▲ ✔

✔ Recommended ▲ Compatible
* CFX384, CFX384 Touch, and MiniOpticon real-time PCR detection systems are factory calibrated for white tubes and white-well plates. White plastics are recommended due to their superior signal-to-noise ratio. Using clear tubes or clear-well plates
on these instruments will require user calibration.

Amplification Reagents and Plastics 34 Visit us on the Web at www.bio-rad.com.

 PCR Plastic Consumables ­
PCR Tubes, Strips, and Caps

0.2 and 0.5 ml Individual PCR Tubes Low-Profile PCR Tube Strips
■■  igh-profile PCR tubes with double-locking caps for a stronger
H ■■ Lower height reduces the potential for condensation
seal during cycling ■■ Designed to allow greater light capture in fluorescence assays www.bio-rad.com/
pcrplastics
■■ Flat, frosted caps are easy to label ■■ Opaque white color option maximizes detection signal
■■  ptions with attached caps for greater convenience and lower
O ■■ Maximum volume: 200 µl
risk of contamination
Flat and Domed Cap Strips for PCR Tubes
High-Profile PCR Tube Strips
■■ Thin-walled tubes for superior heat transfer
and PCR Plates
■■ Ultraclear flat cap strips are ideal for qPCR
■■ 8- and 12-tube strips for use on 48- or 96-well sample blocks
■■  esigned for extremely tight sealing for thermal cycling and storage
D
■■ Available in a variety of colors for easy sample tracking (– 20 and 4˚C)
■■ Maximum volume: 300 µl ■■Flat caps have optimal light transmittance for real-time PCR on PCR
tubes or plates
■■ Great option to save plates if fewer wells are needed during PCR

TLS-0801, clear
TLS-0851, white
Low-Profile Tube Strips without Caps
TCS-0803, ultraclear
Optical Flat 8-Cap Strips

ECT-2000
Strip Cap Tool

TCS-0801, 8-cap, clear

TCS-1201, 12-cap, clear
Domed Cap Strips

Plastics
TBS-0201, 8-tube, clear
TBS-1201, 12-tube, clear
High-Profile Tube Strips without Caps

© 2013 Bio-Rad Laboratories, Inc. Amplification Reagents and Plastics 35

 PCR Plastic Consumables ­
PCR Plates

Multiplate™ 96-Well and 48-Well Unskirted

PCR Plates
www.bio-rad.com/ ■■  ingle-component polypropylene allows low protein binding
S
pcrplastics
and excellent sample retention
■■  igh-profile (20.7 mm) and low-profile (15.5 mm) options allow broad
H
real-time PCR instrument compatibility
■■ White-well option for maximizing real-time PCR detection sensitivity
■■  esigned to allow easy cutting with scissors (when less than a full
D MLP-4801, clear
Multiplate High-Profile 48-Well Unskirted PCR Plates
plate is needed)

iQ™ High-Profile 96-Well Semi-Skirted PCR Plates

■■ Semi-skirt design provides a labeling surface for easy sample tracking
■■ Composition designed to provide stiffness during plate handling
■■  igh-profile (20.7 mm) plate has perforations every 3 columns
H
for convenience (when less than a full plate is needed)

MLL-4801, clear
MLL-4851, white
Multiplate Low-Profile 48-Well Unskirted PCR Plates

MLL-9601, clear
Multiplate Low-Profile 96-Well Unskirted PCR Plates 223-9441
iQ High-Profile 96-Well Semi-Skirted PCR Plates

© 2013 Bio-Rad Laboratories, Inc. Amplification Reagents and Plastics 36

 PCR Plastic Consumables ­
Hard-Shell ® PCR Plates

Hard-Shell Technology Raised rims allow for Rigid skirt ideal

The patented two-component design of Hard-Shell plates is specifically The skirt and deck of a tight sealing for heat sealing
Hard-Shell plate prevent
engineered to withstand the stresses of thermal cycling. Benefits include: warping and shrinkage
www.bio-rad.com/
pcrplastics
■■  uperior stability and flatness allow precise positioning
S
and robotic handling
■■ Sturdy plate design is ideal for heat sealing methods
■■  arp-resistant feature provides durability during automation,
W
high-speed centrifugation, and storage (even –80°C)
■■  ser-readable bar code options for convenient sample tracking
U
in high-throughput settings
■■ Black alphanumeric labeling for easy well identification
■■  ootprint and well spacing designed to match ANSI/SBS
F Thin-wall polypropylene V-shaped wells
enable optimal thermal transfer and
standard dimensions recovery of low-volume samples

■■ Composition helps prevent DNA binding

■■ Polypropylene resin allows superior well-to-well uniformity for reliable
and reproducible real-time qPCR results

3.0

2.5
Relative signal strength

2.0

1.5

1.0

Plastics
0.5

HSP-9955, white shell, white well, bar-coded 0

Clear White Black
Hard-Shell Low-Profile 96-Well Skirted PCR Plates
Other colors and bar code option are available. Enhanced real-time PCR sensitivity.

© 2013 Bio-Rad Laboratories, Inc. Amplification Reagents and Plastics 37

 PCR Plastic Consumables ­
Hard-Shell® PCR Plates

www.bio-rad.com/
pcrplastics

15.51 mm
Low-profile 9.90 mm

Semi-skirt
HSL-9641, green shell, clear well HSP-3821, yellow shell, clear well
Hard-Shell Low-Profile 96-Well Semi-Skirted PCR Plates Hard-Shell 384-Well Skirted PCR Plates

15.51 mm 20.75 mm
Low-profile High-profile

Semi-skirt
Semi-skirt
HSR-9905, clear shell, white well HSS-9665, black shell, white well
Hard-Shell 96-Well 480 PCR Plates Hard-Shell High-Profile 96-Well Semi-Skirted PCR Plates

Other colors and bar code option are available.

Refer to page 34 for an instrument compatibility guide for PCR plates.

Amplification Reagents and Plastics 38 Visit us on the Web at www.bio-rad.com.

 PCR Plastic Consumables ­
PCR Plate Sealing

PX1™ PCR Plate Sealer

The PX1 PCR plate sealer is a semiautomated heat sealer for consistent
and uniform sealing across an entire microplate. Features include: www.bio-rad.com/
pcrplatesealer
■■ Intuitive touch-screen user interface for extreme convenience
and ease of use
■■ Programmable sealing protocols for quick access
■■ Small footprint suitable for crowded laboratory benches
■■  ompatible with a variety of heat sealing films and foils
C
and a wide range of PCR plates

Seals Validated for PX1 PCR Plate Sealer

Optically Clear Heat Seal
■■ 
Ideal for real-time PCR
■■ Excellent optical clarity
■■ Peelable for easy sample retrieval
■■ Compatible with PCR
Permanent Clear Heat Seal
■■ 
Ideal for water bath cycling
■■ Nonpeelable, nonpierceable seal
■■ High solvent resistance
Pierceable Foil Heat Seal
Fully validated for the QX100™ Droplet Digital™ PCR system workflow
■■ 

■■ Easily pierceable with a pipet tip

■■ User friendly — colored stripe clearly identifies sealing surface
■■ Compatible with PCR
Peelable Foil Heat Seal
■■ 
Ideal for low-temperature sample storage

Plastics
■■  an be easily peeled from microplates stored in a –80˚C freezer
C
or in liquid nitrogen
■■ Compatible with PCR

© 2013 Bio-Rad Laboratories, Inc. Amplification Reagents and Plastics 39

 PCR Plastic Consumables ­
PCR Plate Sealing

Microseal ® 'B' Adhesive Seals, Optically Clear Microseal 'F' Foil

■■  trongest adhesive-based optically clear sealing option designed
S ■■  luminum foil allows opaque sealing option for DNA sequencing
A
www.bio-rad.com/ for real-time PCR plates (ABI 3700 DNA analyzer) and sample storage
pcrseals
■■Withstands multiple storage or transport temperatures ■■  cts as a barrier against evaporation in extreme temperatures
A
(–40 to 110°C) (–80 to 105°C)

Microseal 'C' Optical Seals ■■ Pierceable foil for easy sample retrieval
■■  ptically clear adhesive films designed for tight seals even
O
with wells with raised rims
Microseal 'A' Film
■■  nonoptical, nonadhesive sealing option for quick pressure-based
A
■■ Pressure-sensitive adhesive allows easy application during plate sealing sealing of plates
■■ Designed with superior optical properties for real-time PCR ■■  llows easy removal without the risk of aerosol formation,
A
minimizing cross-contamination
■■ Convenient option for standard PCR needs

MSB-1001 MSC-1001 MSR-0001 MSF-1001 MSA-5001

Microseal 'B' Adhesive Seals Microseal 'C' Optical Seals Sealing Roller Microseal 'F' Foil Microseal 'A' Film

Amplification Reagents and Plastics 40 Visit us on the Web at www.bio-rad.com.

 PCR Plastic Consumables ­
Ordering Information

Ordering Information
Catalog # Description
PCR Tubes, Strips, and Caps
Individual PCR Tubes with Attached Caps (0.2 ml)
TFI-0201 PCR Tubes with Flat Caps (0.2 ml), clear, 1,000
TWI-0201 PCR Tubes with Domed Caps (0.2 ml), clear, 1,000
Individual PCR Tubes without Caps (0.2 ml)
TBI-0201 PCR Tubes without Caps (0.2 ml), clear, 1,000
Individual PCR Tubes with Attached Caps (0.5 ml)
TBI-0501 PCR Tubes with Flat Caps (0.5 ml), clear, 1,000 (2 bags of 500)
TBI-0502 PCR Tubes with Flat Caps (0.5 ml), clear, 800 (8 bags of 100)
High-Profile Tube Strips without Caps
TBS-0201 8-Tube Strips without Caps, clear, 120 strips (960 PCR tubes)
TBS-1201 12-Tube Strips without Caps, clear, 100 strips (1,200 PCR tubes)
Low-Profile 8-Tube Strips without Caps
TLS-0801 Low-Profile 8-Tube Strips without Caps, clear, 120 (960 PCR tubes)
TLS-0851 Low-Profile 8-Tube Strips without Caps, white, 120 (960 PCR tubes)
Domed Cap Strips
TCS-0801 Domed 8-Cap Strips, for PCR tubes and plates, clear, 120
TCS-1201 Domed 12-Cap Strips, for PCR tubes and plates, clear, 200
Optical Flat Cap Strips
TCS-0803 Optical Flat 8-Cap Strips, for PCR tubes and plates, ultraclear, 120
High-Profile Tube Strips with Domed Cap Strips
TBC-0802 8-Tube Strips and Domed Cap Strips, clear, 20 bags of 12 x 8-tube strips and 12 x 8-cap strips (1,920 PCR tubes and 1,920 caps)
TBC-1202 12-Tube Strips and Domed Cap Strips, clear, 20 bags of 8 x 12-tube strips and 8 x 12-cap strips (1,920 PCR tubes and 1,920 caps)
Capping Tools and Racks
TRC-9601 PCR Tube Rack, ANSI/SBS standard, white, 10
TRC-0501 96-Place Racks, with covers, for PCR tubes and unskirted and semi-skirted microplates, assorted colors, 5
ECT-1000 Easy Cap Tool, ensures tight seal for 0.2 ml PCR tubes or 96-well microplates
ECT-2000 Strip Cap Tool, for sealing 8- and 12-cap strips on PCR plates or tubes
PCR Plates
Multiplate 48-Well PCR Plates
MLP-4801 Multiplate High-Profile 48-Well Unskirted PCR Plates, clear, 50 plates
MLL-4801 Multiplate Low-Profile 48-Well Unskirted PCR Plates, clear, 50 plates
MLL-4851 Multiplate Low-Profile 48-Well Unskirted PCR Plates, white, 50 plates
Multiplate High-Profile 96-Well Unskirted PCR Plates
MLP-9601 Multiplate High-Profile 96-Well Unskirted PCR Plates, clear, 25 plates
MLP-9651 Multiplate High-Profile 96-Well Unskirted PCR Plates, white, 25 plates
MLP-9631 Multiplate High-Profile 96-Well Unskirted PCR Plates, blue, 25 plates

Plastics
Multiplate Low-Profile 96-Well Unskirted PCR Plates
MLL-9601 Multiplate Low-Profile 96-Well Unskirted PCR Plates, clear, 25 plates
MLL-9651 Multiplate Low-Profile 96-Well Unskirted PCR Plates, white, 25 plates
iQ High-Profile 96-Well Semi-Skirted PCR Plates
223-9441 iQ High-Profile 96-Well Semi-Skirted PCR Plates, 25 plates

© 2013 Bio-Rad Laboratories, Inc. Amplification Reagents and Plastics 41

 PCR Plastic Consumables ­
Ordering Information

Ordering Information
Description Clear Well White Well Black Well
Hard-Shell Plates
Hard-Shell Low-Profile 96-Well Skirted PCR Plates
White shell, 50 HSP-9601 HSP-9655 —
Red shell, 50 HSP-9611 — —
Yellow shell, 50 HSP-9621 — —
Blue shell, 50 HSP-9631 HSP-9635 —
Green shell, 50 HSP-9641 HSP-9645 —
Black shell, 50 HSP-9661 HSP-9665 HSP-9666
White shell, bar-coded, 50 HSP-9901 HSP-9955 —
Hard-Shell High-Profile 96-Well Semi-Skirted PCR Plates
Clear shell, 25 HSS-9601 — —
Green shell, 25 HSS-9641 — —
Black shell, 25 — HSS-9665 —
Clear shell, bar-coded, 25 HSS-9901 — —
Hard-Shell Low-Profile 96-Well Semi-Skirted PCR Plates
Clear shell, 25 HSL-9601 HSL-9605 —
Green shell, 25 HSL-9641 HSL-9645 —
Clear shell, bar-coded, 25 HSL-9901 HSL-9905 —

25 Plates 100 Plates, 100 Microseal 'C' Seals
Hard-Shell 96-Well 480 PCR Plates with Bar Code on Row A Side
Clear shell, white well HSR-9905 HSR-9905K
Clear shell, clear well HSR-9901 HSR-9901K

Amplification Reagents and Plastics 42 Visit us on the Web at www.bio-rad.com.

PCR Plastic Consumables ­
Ordering Information

Ordering Information
Description Clear Well White Well Black Well
Hard-Shell Plates cont.
Hard-Shell 384-Well Standard PCR Plates
Clear shell, 50 HSP-3801 HSP-3805 —
Red shell, 50 HSP-3811 — —
Yellow shell, 50 HSP-3821 — —
Blue shell, 50 HSP-3831 — —
Green shell, 50 HSP-3841 — —
Black shell, 50 — HSP-3865 HSP-3866
Clear shell, bar-coded, 50 HSP-3901 HSP-3905 —

50 Plates 100 Plates, 100 Microseal 'C' Seals

Hard-Shell 384-Well 480 PCR Plates with Bar Code on Row A Side
Clear shell, white well HSR-4805 HSR-4805K
Clear shell, clear well HSR-4801 HSR-4801K
Catalog # Description
PCR Plate Sealing
181-4000 PX1 PCR Plate Sealer, includes heat sealing instrument, 96-well/384-well plate support block, sealing frame, power cord
181-4030 Optically Clear Heat Seal, 100 seals
181-4035 Permanent Clear Heat Seal, 100 seals
181-4040 Pierceable Foil Heat Seal, 100 seals
181-4045 Peelable Foil Heat Seal, 100 seals
MSA-5001 Microseal 'A' Film, 50 seals
MSB-1001 Microseal 'B' Adhesive Seals, optically clear, 100 seals
MSC-1001 Microseal 'C' Optical Seals, 100 seals
MSF-1001 Microseal 'F' Foil, 100 seals
MSR-0001 Sealing Roller, for film seals
ADR-3296 Optical Compression Pad, for improved film sealing of 96-well plates in DNA Engine Opticon 2 and Chromo4 systems
ADR-5001 Pressure Pad, uniformly distributes lid pressure for sealing film
MSO-1001 Optical Film Sealing Kit, for 96-well plates, includes optical compression pad, 100 Microseal 'B' clear adhesive seals
223-9444 Optical Sealing Tape, 100 sheets
223-9442 96-Well PCR Plate Sealing Mats, 5

Plastics
 Reference
Bustin SA et al. (2009). The MIQE guidelines: Minimum information for publication of quantitative real-time PCR experiments. Clin Chem 55, 611–622.

Cy is a trademark of GE Healthcare group companies. Eppendorf and Mastercycler are trademarks of Eppendorf AG. EvaGreen is a trademark of Biotium, Inc. Bio-Rad Laboratories, Inc. is
licensed by Biotium, Inc. to sell reagents containing EvaGreen dye for use in real-time PCR, for research purposes only. FAM, ROX, StepOne, StepOnePlus, and Veriti are trademarks of Applera
Corporation. HRM and Rotor-Gene are trademarks of QIAGEN GmbH. LightCycler is a trademark of Roche Diagnostics GmbH. LightScanner is a trademark of Idaho Technology Inc. Mx,
Mx3000P, Mx3005P, and Mx4000 are trademarks of Stratagene Corporation. SYBR and ViiA are trademarks of Life Technologies Corporation. Bio-Rad Laboratories, Inc. is licensed by
Life Technologies Corporation to sell reagents containing SYBR Green I for use in real-time PCR, for research purposes only. Texas Red is a trademark of Invitrogen Corporation.

Notice regarding Bio-Rad thermal cyclers and real-time systems:

Purchase of this instrument conveys a limited non-transferable immunity from suit for the purchaser’s own internal research and development and for use in human in vitro diagnostics and all other
applied fields under U.S. Patent Number 5,475,610 (Claims 1, 44, 158, 160–163, and 167 only), or corresponding claims in its non-U.S. counterpart, owned by Applera Corporation. No right is conveyed
expressly, by implication, or by estoppel under any other patent claim, such as claims to apparatus, reagents, kits, or methods such as 5' nuclease methods. Further information on purchasing licenses
may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.

Bio-Rad’s real-time thermal cyclers are licensed real-time thermal cyclers under Applera’s U.S. Patent Number 6,814,934 B1 for use in research, human in vitro diagnostics, and all other fields
except veterinary diagnostics.

Bio-Rad’s thermal cyclers and real-time thermal cyclers are covered by one or more of the following U.S. patents or their foreign counterparts owned by Eppendorf AG: U.S. Patent Numbers
6,767,512 and 7,074,367.

Practice of the patented 5' Nuclease Process requires a license from Applied Biosystems. The purchase of these products includes an immunity from suit under patents specified in the product insert
to use only the amount purchased for the purchaser’s own internal research when used with the separate purchase of Licensed Probe. No other patent rights are conveyed expressly, by implication,
or by estoppel. Further information on purchasing licenses may be obtained from the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.

Hard-Shell plates are covered by one or more of the following U.S. patents or their foreign counterparts owned by Eppendorf AG: U.S. Patent Numbers 7,347,977; 6,340,589; and 6,528,302.

Bio-Rad
Laboratories, Inc.

Life Science Web site www.bio-rad.com USA 800 424 6723 Australia 61 2 9914 2800 Austria 01 877 89 01 Belgium 09 385 55 11 Brazil 55 11 5044 5699 Canada 905 364 3435 China 86 21 6169 8500 Czech Republic 420 241 430 532
Denmark 44 52 10 00 Finland 09 804 22 00 France 01 47 95 69 65 Germany 089 31 884 0 Greece 30 210 9532 220 Hong Kong 852 2789 3300 Hungary 36 1 459 6100 India 91 124 4029300 Israel 03 963 6050
Group Italy 39 02 216091 Japan 03 6361 7000 Korea 82 2 3473 4460 Mexico 52 555 488 7670 The Netherlands 0318 540666 New Zealand 64 9 415 2280 Norway 23 38 41 30 Poland 48 22 331 99 99 Portugal 351 21 472 7700
Russia 7 495 721 14 04 Singapore 65 6415 3188 South Africa 27 861 246 723 Spain 34 91 590 5200 Sweden 08 555 12700 Switzerland 026 674 55 05 Taiwan 886 2 2578 7189 Thailand 800 88 22 88
United Kingdom 020 8328 2000

Bulletin 6090 Rev D US/EG 12-1779 0213 Sig 1212