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Amplification: Consumables

Amplification: Consumables Amplification Reagents and Plastics
Amplification: Consumables Amplification Reagents and Plastics

Amplification Reagents and Plastics

Amplification: Consumables Amplification Reagents and Plastics
Amplification: Consumables Amplification Reagents and Plastics
Amplification: Consumables Amplification Reagents and Plastics
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 ■■ Assay specificity ■■ Inhibitors in sample ■■ Reproducibility of thermal cycling conditions and instrument compatibility

PCR Plastic Consumables ■■ Instrument compatibility ■■ Optimum performance ■■ Automation friendly ■■ Potential source of contamination and inhibition

RNA Isolation ■■ Kits are designed and formulated to assist in the isolation of highly
RNA Isolation ■■ Kits are designed and formulated to assist in the isolation of highly
RNA Isolation ■■ Kits are designed and formulated to assist in the isolation of highly
RNA Isolation ■■ Kits are designed and formulated to assist in the isolation of highly
RNA Isolation ■■ Kits are designed and formulated to assist in the isolation of highly
RNA Isolation ■■ Kits are designed and formulated to assist in the isolation of highly
RNA Isolation ■■ Kits are designed and formulated to assist in the isolation of highly
RNA Isolation ■■ Kits are designed and formulated to assist in the isolation of highly
RNA Isolation ■■ Kits are designed and formulated to assist in the isolation of highly

RNA Isolation

■■ Kits are designed and formulated to assist in the isolation of highly pure and intact RNA from different starting materials

■■ RNA is compatible with a variety of downstream applications

– Real-time qPCR

– Northern blotting

– Microarray analysis

– cDNA library construction

■■ DNase treatment ensures genomic DNA removal

RNA Isolation

RNA Isolation
RNA Isolation

Aurum Total RNA Fatty and Fibrous Tissue Kit

■■ PCR-ready RNA in less than 60 min

■■ PureZOL efficiently lyses cells and tissues, deproteinates RNA, and inactivates endogenous nucleases in a single step

■■ High yield of intact total RNA from difficult-to-disrupt samples, including plant and animal tissues

■■ Well suited for fungal samples that are rich in RNases

■■ RNase-free reagents and plastic consumables ensure the integrity of isolated RNA

■■ Kit includes DNase I for removal of genomic DNA contamination

■■ Easy-to-use spin or vacuum protocol

Aurum Total RNA Mini Kit

■■ PCR-ready RNA in less than 60 min

■■ Guanidine isothiocyanate and β-mercaptoethanol efficiently lyse samples and quickly inactivate RNases

■■ High yield of intact total RNA from a wide range of starting materials, including cultured cells, bacteria, and yeast, as well as plant and animal tissues

■■ RNase-free reagents and plastic consumables ensure the integrity of isolated RNA

Aurum Total RNA 96 Kit

■■ High-throughput total RNA isolation in less than 60 min

■■ High yield of intact total RNA from a wide range of starting materials, including cultured cells, bacteria, and yeast, as well as plant and animal tissues

■■ Guanidine isothiocyanate and β-mercaptoethanol efficiently lyse samples and quickly inactivate RNases

■■ RNase-free reagents and plastic consumables ensure the integrity of isolated RNA

■■ Kit includes DNase I for removal of genomic DNA contamination

■■ Compatible with Aurum vacuum manifold

732-6800, 2 x 96-well preps Aurum Total RNA 96 Kit
732-6800, 2 x 96-well preps
Aurum Total RNA 96 Kit
732-6800, 2 x 96-well preps Aurum Total RNA 96 Kit ■■ Kit includes DNase I for

■■ Kit includes DNase I for removal of genomic DNA contamination

■■ Easy-to-use spin or vacuum protocol

vacuum protocol For more information, request bulletin 2920. 732-6830, 50 preps Aurum Total RNA Fatty and

732-6830, 50 preps Aurum Total RNA Fatty and Fibrous Tissue Kit

732-6820, 50 preps Aurum Total RNA Mini Kit

RNA Isolation
RNA Isolation

iScript RT-qPCR Sample Preparation Reagent

■■ Reagent stabilizes RNA and removes genomic DNA in less than 10 min

■■ Suitable for adherent or suspension animal cells

■■ RT-qPCR is directly enabled from cells without RNA purification when combined with an iScript reverse transcription kit and real-time supermix

■■ Reagent allows multiplex real-time detection of up to 4 targets from as few as 10 cells

■■ Ideal for rapid, high-throughput gene expression analysis

4 10 3 10 2 10 RFU
4
10
3
10
2
10
RFU

0

10

20

Cycles

30

40

PureZOL RNA Isolation Reagent

■■ Single-solution format permits recovery of RNA from small quantities of tissues or cells, making it ideally suited for gene expression studies

■■ Efficient RNA purification from cultured cells, yeast, viruses, and bacteria, as well as plant and animal tissues

■■ PureZOL efficiently lyses cells and tissues, deproteinates RNA, and inactivates endogenous nucleases in a single step

■■ Scalable starting sample amount

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

Aurum Vacuum Manifold

■■ Vacuum-mediated nucleic acid purification platform

■■ Versatile manifold format adaptable for 96-well plate or up to 18 spin columns

■■ Manifold ensures fast, high-quality sample preparation while maintaining the simplicity of vacuum processing

■■

Unique vacuum regulator design allows for complete control of negative pressure

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.

PCR detection system. RFU, relative fluorescence units. 170-8899, 5 x 10 ml iScript RT-qPCR Sample Preparation

170-8899, 5 x 10 ml iScript RT-qPCR Sample Preparation Reagent

5 x 10 ml iScript RT-qPCR Sample Preparation Reagent 732-6890, 100 ml PureZOL RNA Isolation Reagent

732-6890, 100 ml PureZOL RNA Isolation Reagent

Reagent 732-6890, 100 ml PureZOL RNA Isolation Reagent 732-6470, 1 unit Aurum Vacuum Manifold Amplification

732-6470, 1 unit Aurum Vacuum Manifold

RNA Isolation

RNA Isolation Selection Guide
RNA Isolation
Selection Guide
 

Aurum Total RNA Kits

 

PureZOL RNA Isolation Reagent

 

Mini

Fatty and Fibrous Tissue

 

96

Format

Mini column Filtration (vacuum or spin)

Mini column Filtration (vacuum or spin)

96-well plate Filtration (vacuum or spin)

Single solution

Maximum starting material amounts Cultured cells Bacterial cells Yeast cells Hard animal tissue Soft to moderately hard animal tissue Plant tissue

organic extraction

2

x 10 6

1

x

10 7

1

x 10 6

1

x

10 7

2.4 x 10 9

2.4 x 10 9

8

x 10 8

2.4 x 10 9

3

x 10 7

3

x

10 7

2

x 10 7

3

x

10 7

20

mg

100

mg

100

mg

40

mg

100

mg

100

mg

40

mg

100

mg

100

mg

Isolation method

Silica membrane

Lysis with PureZOL reagent, purification on silica membrane

Silica membrane

Organic extraction

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 Aurum Total RNA Fatty and Fibrous Tissue Module Aurum Total RNA Mini Kit Aurum Total RNA 96 Kit iScript RT-qPCR Sample Preparation Reagent, 100 reactions

170-8899

iScript RT-qPCR Sample Preparation Reagent, 500 reactions PureZOL RNA Isolation Reagent, 50 ml PureZOL RNA Isolation Reagent, 100 ml Aurum Vacuum Manifold

732-6870*

732-6880

732-6820

732-6890

732-6800

732-6470

170-8898

 

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

 
Reagents — Reverse Transcription ■■ Formulated for efficient reverse transcription across a broad linear dynamic
Reagents — Reverse Transcription ■■ Formulated for efficient reverse transcription across a broad linear dynamic
Reagents — Reverse Transcription ■■ Formulated for efficient reverse transcription across a broad linear dynamic
Reagents — Reverse Transcription ■■ Formulated for efficient reverse transcription across a broad linear dynamic
Reagents — Reverse Transcription ■■ Formulated for efficient reverse transcription across a broad linear dynamic
Reagents — Reverse Transcription ■■ Formulated for efficient reverse transcription across a broad linear dynamic
Reagents — Reverse Transcription ■■ Formulated for efficient reverse transcription across a broad linear dynamic

Reagents — Reverse Transcription

■■ Formulated for efficient reverse transcription across a broad linear dynamic range

■■ Potent RNase A inhibitors protect RNA during setup and reverse transcription

■■ Flexible input RNA capacity to suit different experimental needs

■■ Optimized for gene expression analysis using real-time qPCR

Reagents

Reagents Reverse Transcription — iScript ™ Kit Selector
Reagents
Reverse Transcription — iScript ™ Kit Selector
Reduce pipetting variability
Reduce
pipetting
variability
Maximize data from single 20 μl reaction
Maximize data
from single
20 μl reaction
Fast and easy to use
Fast and
easy to use
Select my own primers
Select my
own primers
One-step RT-qPCR with SYBR ® Green
One-step
RT-qPCR with
SYBR ® Green
One-step RT-qPCR for probes
One-step
RT-qPCR for
probes
iScript reverse iScript advanced cDNA synthesis kit for RT-qPCR iScript cDNA transcription supermix synthesis kit
iScript reverse
iScript advanced
cDNA synthesis kit
for RT-qPCR
iScript cDNA
transcription supermix
synthesis kit
iScript Select
cDNA synthesis kit
for RT-qPCR
iScript ™ one-step
RT-PCR kit
with SYBR ® Green
iScript one-step
RT-PCR kit for probes
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
transcriptase
transcriptase
transcriptase
iScript reverse
transcriptase
(for one-step RT-PCR)
iScript reverse
transcriptase
(for one-step RT-PCR)
5x iScript
5x iScript
advanced
RT supermix
(dNTPs, oligo[dT],
random
primers,
buffer components,
and iScript reverse
transcriptase)
reaction mix
(dNTPs, oligo[dT],
random primers, and
buffer components)
5x iScript reaction mix
(dNTPs, oligo[dT],
random primers, and
buffer components)
5x iScript reaction mix
(dNTPs and
buffer components)
2x SYBR ® Green
RT-PCR reaction mix
(dNTPs, iTaq ™ DNA
polymerase,
uorescein,
SYBR ® Green I dye,
and stabilizers)
2x probes RT-PCR
reaction mix (dNTPs,
iTaq DNA polymerase,
and stabilizers)
Oligo(dT),
random primers, and
Forward and
Forward and
reverse
primers
reverse
primers
gene-speci c primer
(GSP) enhancer
for target gene
(not included)
solution
(3 vials)
and probe for
target gene
(not included)
cDNA ready in
40 min for qPCR
cDNA ready in 35 min
for qPCR
cDNA ready in 40 min
for qPCR
cDNA ready in
40–90 min for qPCR
RT-qPCR data in
60–90 min
RT-qPCR data
in 60 min
SensitivityKit
Contents
Reagents Two-Step Reverse Transcription Reagents
Reagents
Two-Step Reverse Transcription Reagents

Cq

iScript Advanced cDNA Synthesis Kit for RT-qPCR

■■ Increased qPCR data throughput and cost effectiveness from a single 20 µl reverse transcription (RT) reaction

■■ Superior sensitivity and broad linear dynamic range for RT (7.5 µg–100 fg)

■■ 2-tube kit (5x iScript reaction mix and iScript reverse transcriptase) for ease of use and reduced reaction setup time

■■ Optimized blend of oligo(dT) and random primers ensures complete and unbiased RNA sequence representation

■■ RNase H+ MMLV reverse transcriptase (preblended with RNase inhibitor) delivers high sensitivity for real-time RT-qPCR and eliminates additional RNase H+ step

■■ Potent blend of RNaseA inhibitor protects RNA during setup and RT

■■ Short 35 min protocol allows fast qPCR data generation

iScript Reverse Transcription Supermix for RT-qPCR

■■ 1-tube format for simple and fast setup, and reduced pipetting variability

■■

Liquid format at –20°C offers superior stability and eliminates freeze/thaw cycle

■■ Superior sensitivity and broad linear dynamic range for RT (1 µg–100 fg)

■■ Optimized blend of oligo(dT) and random primers ensures complete and unbiased RNA sequence representation

■■ RNase H+ MMLV reverse transcriptase (preblended with RNase inhibitor) delivers high sensitivity for real-time RT-qPCR and eliminates additional RNase H+ step

■■ Potent blend of RNaseA inhibitor protects RNA during setup and RT

■■ Short 40 min protocol allows fast qPCR data generation

35

30

25

20

15

10 3 10 2 0 10 20 30 40 Cycles RFU
10
3
10
2
0
10
20
30
40
Cycles
RFU

–2

–1

0

1

log starting quantity

2

3

4

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 R 2 = 0.999, efficiency = 90.7%, slope = –3.57. Cq, quantification cycle; RFU, relative fluorescence units.

10 4 — 1 µg — 100 ng — 10 ng — 1 ng —
10
4
— 1 µg
— 100 ng
— 10 ng
— 1 ng
— 100 pg
— 10 pg
— 1 pg
10
3
RFU

0

10

20

Cycles

30

40

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 β-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.

RNA Average Cq SD CV, % 10 4 100 ng 21.35 0.123 0.576 100 pg
RNA
Average Cq
SD
CV, %
10
4
100 ng
21.35
0.123
0.576
100 pg
31.56
0.147
0.465
10
3
10
2
RFU

10

20

Cycles

30

40

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.

Reagents Two-Step Reverse Transcription Reagents
Reagents
Two-Step Reverse Transcription Reagents
Reagents Two-Step Reverse Transcription Reagents

iScript cDNA Synthesis Kit

■■ 2-tube kit (5x iScript reaction mix and iScript reverse transcriptase) for ease of use and reduced reaction setup time

■■ Superior sensitivity and broad linear dynamic range for RT (1 µg–100 fg)

■■ Optimized blend of oligo(dT) and random primers ensures complete and unbiased RNA sequence representation

■■ RNase H+ MMLV reverse transcriptase (preblended with RNase inhibitor) delivers high sensitivity for real-time RT-qPCR and eliminates additional RNase H+ step

■■ Potent blend of RNaseA inhibitor protects RNA during setup and RT

■■ Short 40 min protocol allows fast qPCR data generation

iScript Select cDNA Synthesis Kit

■■ 5-tube kit (random primers, oligo[dT], 5x iScript Select reaction mix, iScript reverse transcriptase, and gene-specific primer enhancer solution)

■■ Choice of priming strategy

■■ Reliable synthesis of long cDNA >6 kb in length

■■ Superior sensitivity and broad linear dynamic range for RT (1 µg–1 pg)

10 3 — 1 µg — 100 ng — 10 ng — 1 ng —
10
3
— 1 µg
— 100 ng
— 10 ng
— 1 ng
— 100 pg
— 10 pg
10
2
— 1 pg
10
RFU
RFU
10 4 10 3 10 2 RFU
10
4
10
3
10
2
RFU
Reagents 10 4 10 3 10 2
Reagents
10
4
10
3
10
2

0

5

10

15

20

25

30

35

40

45

0

10

20

30

40

0

2

4

6

 

Cycles

Cycles

8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 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 r 2 = 0.998, efficiency = 96.5%. RFU, relative fluorescence units.

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.

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 β-actin gene with iQ SYBR ® Green supermix. Standard curve r = 1.000, efficiency = 92.2%. RFU, relative fluorescence units.

Reagents One-Step RT-qPCR Reagents
Reagents
One-Step RT-qPCR Reagents

iScript One-Step RT-PCR Kit with SYBR ® Green

■■ For use on a broad range of real-time PCR instruments

■■ Extremely sensitive detection (100 ng–1 pg) of input RNA

iScript One-Step RT-PCR Kit for Probes

■■ For use with all types of hybridization probes, including dual-labeled oligonucleotide probes, FRET probes, and molecular beacons

■■ Extremely sensitive detection (1 µg–1 pg) of input RNA

10 4 10 3 10 2 PCR baseline-subtracted curve fit, RFU
10
4
10
3
10
2
PCR baseline-subtracted curve fit, RFU

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 Cycles

iScript one-step RT-PCR kit with SYBR ® Green provides high reproducibility and sensitivity across a broad range of concentrations. Reactions were performed in triplicate, along with no-template controls, using GAPDH primers and 100 ng–100 fg total HeLa RNA. Reactions were carried out on the iCycler iQ ® real-time PCR detection system. Standard curve r = 1.000, efficiency = 95%, slope = –3.47. RFU, relative fluorescence units.

Benefits of iScript one-step kits:

■■ Provide powerful combination of iScript RNase H+ reverse transcriptase and antibody-mediated hot-start iTaq DNA polymerase

■■ Are ideal for rapid, high-throughput gene expression analysis

■■ Perform cDNA synthesis and qPCR in 1 tube, minimizing handling and contamination risk

10 3 10 2 10 PCR baseline-subtracted curve fit, RFU
10
3
10
2
10
PCR baseline-subtracted curve fit, RFU

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

iScript one-step RT-PCR kit for probes delivers unparalleled results over an extremely wide dynamic range. RNA (1 µg–100 fg) isolated from HeLa cells using the Aurum total RNA kit was reverse transcribed and amplified using primers to β-actin and a FAM-labeled detection probe. Each dilution was performed in triplicate and RT-PCR was carried out on the 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 iScript Advanced cDNA Synthesis Kit for RT-qPCR, 250 x 20 μl reactions iScript cDNA Synthesis Kit, 25 x 20 μl reactions iScript cDNA Synthesis Kit, 100 x 20 μl reactions iScript Reverse Transcription Supermix for RT-qPCR, 25 x 20 μl reactions iScript Reverse Transcription Supermix for RT-qPCR, 100 x 20 μl reactions iScript Select cDNA Synthesis Kit, 25 x 20 μl reactions iScript Select cDNA Synthesis Kit, 100 x 20 μl reactions

170-8892

iScript One-Step RT-PCR Kit with SYBR Green, 50 x 50 μl reactions iScript One-Step RT-PCR Kit with SYBR Green, 200 x 50 μl reactions iScript One-Step RT-PCR Kit for Probes, 50 x 50 μl reactions iScript One-Step RT-PCR Kit for Probes, 200 x 50 μl reactions

170-8843

170-8893

170-8890

170-8894

170-8891

170-8895

170-8840

 

170-8841

170-8896

170-8897

Reagents

Reagents Reagents — Real-Time qPCR ■■ Patented Sso7d fusion enzyme technology delivers higher processivity and
Reagents Reagents — Real-Time qPCR ■■ Patented Sso7d fusion enzyme technology delivers higher processivity and

Reagents — Real-Time qPCR

■■ Patented Sso7d fusion enzyme technology delivers higher processivity and inhibitor tolerance

■■ Antibody-mediated hot-start technology enables instant polymerase activation and superior specificity

■■ Choice of fast, standard, or universal cycling conditions

■■ Formulated for optimal performance on a variety of real-time instruments

Reagents Real-Time qPCR Supermixes
Reagents
Real-Time qPCR Supermixes
Reagents Real-Time qPCR Supermixes Property SsoAdvanced ™ iTaq ™ Universal Supermixes iQ ™
Reagents Real-Time qPCR Supermixes Property SsoAdvanced ™ iTaq ™ Universal Supermixes iQ ™
Reagents Real-Time qPCR Supermixes Property SsoAdvanced ™ iTaq ™ Universal Supermixes iQ ™
Reagents Real-Time qPCR Supermixes Property SsoAdvanced ™ iTaq ™ Universal Supermixes iQ ™

Property

SsoAdvanced

iTaq Universal Supermixes

iQ Supermixes

Application-Specific Kits and Reagents

Supermixes

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

• • •

Reagents

Reagents Real-Time qPCR Supermixes
Reagents
Real-Time qPCR Supermixes

SsoAdvanced Supermixes

■■ Superior performance even from compromised samples

■■ Tolerance for a broad range of reaction conditions and difficult amplicons

■■ 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

■■ 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

■■ Superior high resolution melt (HRM) data for single nucleotide polymorphism (SNP) detection

■■ Accurate methylation detection

■■ Novel quantitative chromatin structure information using qPCR

Reagents SsoAdvanced ™ Supermixes
Reagents
SsoAdvanced ™ Supermixes

SsoAdvanced SYBR ® Green Supermix*

■■ Novel Sso7d fusion polymerase enables increased resistance to PCR inhibitors and higher processivity for dye-based real-time qPCR

■■ Robust formulation delivers maximum efficiency, sensitivity, and reproducibility across a broad range of standard and fast cycling conditions

■■ Antibody-mediated hot-start technology and optimized buffer allow for instant polymerase activation and rapid polymerization kinetics to enable fast PCR

■■ Advanced formulation tolerates a broad range of reaction conditions, primer concentrations, and temperature ranges

* SsoAdvanced universal SYBR ® Green supermix will be available soon and it will be compatible with any real-time PCR instrument.

Sso7d Polymerase
Sso7d
Polymerase

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.

10 5 38 36 34 32 30 10 4 28 26 –12 –11 –10 –9
10
5
38
36
34
32
30
10
4
28
26
–12
–11
–10
–9
–8
log starting quantity
10
3
10
2
RFU
Cq

0

10

20

Cycles

30

40

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 R 2 = 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.

10 4 700 600 500 400 10 3 300 200 100 0 65 70 75
10
4
700
600
500
400
10
3
300
200
100
0
65
70
75
80
85
90
95
Temperature, °C
10
2
10
RFU
–d(RFU)/dT

0

10

20

Cycles

30

40

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.

10 5 10 4 18 19 20 21 22 Cycles 10 3 10 2 RFU
10
5
10
4
18
19
20
21
22
Cycles
10
3
10
2
RFU
RFU

0

10

20

Cycles

30

40

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 R 2 = 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.

Reagents SsoAdvanced ™ Supermixes
Reagents
SsoAdvanced ™ Supermixes

SsoFast Probes Supermix

■■ Novel Sso7d fusion polymerase enables increased resistance to PCR inhibitors and higher processivity for probe-based real-time qPCR

■■ Robust formulation allows simultaneous detection of up to 2 different gene targets using fluorogenic probes with maximum efficiency, sensitivity, and reproducibility across a broad range of standard and fast cycling conditions

■■ Antibody-mediated hot-start technology and optimized buffer allow for instant polymerase activation and rapid polymerization kinetics to enable fast PCR

Sso7d Polymerase
Sso7d
Polymerase

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.

10 5 10 4 10 3 10 2 RFU
10
5
10
4
10
3
10
2
RFU

0

10

20

Cycles

30

40

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. () HEX-labeled GAPDH probe duplex reaction; () Texas Red–labeled IL-2 probe duplex reaction; () HEX-labeled GAPDH probe simplex reaction; () Texas Red–labeled IL-2 probe simplex reaction. Total qPCR run time = 38 min. RFU, relative fluorescence units.

10 4 10 3 log starting quantity 10 2 10 1 RFU Cq
10
4
10
3
log starting quantity
10
2
10
1
RFU
Cq

0

10

20

Cycles

30

40

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%, R 2 = 0.997. Inset shows the standard curve for the various dilutions. Total qPCR run time = 50 min. Cq, quantification cycle; RFU, relative fluorescence units.

Plate #

Start Time,

PCR

R

2

hr

Efficiency, %

 

1

0

97.0

0.999

2

14.0

97.6

0.999

3

24.5

95.9

0.999

4

39.5

97.3

0.999

5

48.0

95.9

0.999

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.

Reagents iTaq ™ Universal Supermixes
Reagents
iTaq ™ Universal Supermixes

iTaq Universal SYBR ® Green Supermix

■■ Advanced 2x ready-to-use supermix, formulated to deliver robust qPCR results with superior sensitivity, efficiency, and specificity

■■ Optimized buffer allows consistent results using both standard and fast cycling protocols

■■ Antibody-mediated iTaq DNA polymerase enables fast activation and superior specificity in qPCR

■■ Formulation developed for optimal results on any real-time PCR instrument

10 4 10 3 17 18 19 20 21 22 23 24 25 26 Cycles
10
4
10
3
17
18
19
20
21
22
23
24
25
26
Cycles
10
3
10
2
RFU
RFU

0

5

10

15

20

Cycles

25

30

35

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 β-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.

10 4 10 3 10 2 RFU
10
4
10
3
10
2
RFU

0

10

20

Cycles

30

40

iTaq universal SYBR ® Green supermix provides reliable gene expression data. The human β-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 R 2 = 0.999, ß-actin efficiency = 92.6%, slope = –3.51. RFU, relative fluorescence units.

10 5 10 4 10 3 10 2 10 RFU
10
5
10
4
10
3
10
2
10
RFU

0

10

20

Cycles

30

40

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 R 2 = 0.993, slope = –3.14. RFU, relative fluorescence units.

Reagents iTaq ™ Universal Supermixes
Reagents
iTaq ™ Universal Supermixes

iTaq Universal Probes Supermix

■■ Advanced 2x ready-to-use supermix, formulated to deliver robust qPCR results with superior sensitivity, efficiency, and specificity

■■ Optimized buffer allows consistent results for simplex and duplex reactions using both standard and fast cycling protocols

■■ Antibody-mediated iTaq DNA polymerase enables fast activation and superior specificity in qPCR

■■ Formulation developed for optimal results on any real-time PCR instrument

10 4 10 3 10 2 RFU
10
4
10
3
10
2
RFU
10 4 10 2 10 3 18 20 22 24 26 Cycles 10 2 RFU
10
4
10
2
10
3
18
20
22
24
26
Cycles
10
2
RFU
RFU
10 4 10 3 10 2 RFU
10
4
10
3
10
2
RFU

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 β-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 β-actin efficiency of 94.3% over 6 orders of linear dynamic range. Standard curve R 2 = 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 β-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-1β 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-1β gene even with very low cDNA inputs. RFU, relative fluorescence units.

Reagents iQ ™ Supermixes
Reagents
iQ ™ Supermixes

iQ SYBR ® Green Supermix

■■ Analysis of low-, medium-, and high-abundance target genes with superior sensitivity and efficiency

■■ Formulated for maximum SYBR ® Green I stability and performance in a wide variety of real-time PCR instruments

■■ Antibody-mediated hot-start polymerase for quick activation and increased specificity

10 3 10 2 PCR baseline-subtracted curve fit, RFU
10
3
10
2
PCR baseline-subtracted curve fit, RFU

0

5

10

15

20

25

Cycles

30

35

40

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-1β 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.

10 4 10 3 PCR baseline-subtracted curve fit, RFU
10
4
10
3
PCR baseline-subtracted curve fit, RFU

0

5

10

15

20

25

Cycles

30

35

40

44

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 β-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.

Reagents

Reagents iQ ™ Supermixes
Reagents
iQ ™ Supermixes

iQ Supermix

■■ Maximum efficiency and sensitivity for qPCR using fluorogenic probes

■■ Reliable amplification over a wide dynamic range of human genomic and plasmid DNA concentrations

■■ Contains antibody-mediated hot-start iTaq DNA polymerase for quick activation and increased specificity

10 3 10 2 RFU
10
3
10
2
RFU

0

5

10

15

20

25

Cycles

30

35

40

45

50

iQ supermix provides sensitive real-time detection over 8 orders of magnitude. Tenfold dilutions of a plasmid containing 10 9 –10 copies of the α-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

■■ Robust supermix formulated for sensitive and efficient multiplex qPCR

■■ Reliable quantification of up to 4 targets (expression levels can vary up to 10 6 -fold between target genes) or up to 5 targets

■■ Linearity over 6 orders of magnitude of input cDNA and 4 orders of magnitude of input genomic DNA

■■ Suitable for a wide variety of applications, including gene expression analysis, SNP genotyping, SNP analysis, GMO detection, and viral load detection

10 3 10 2 RFU
10
3
10
2
RFU

0

5

10

15

20

25

30

35

40

45

 

Cycles

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 β-actin probe (), Cy5-labeled α-tubulin probe (), HEX-labeled GAPDH probe (), TAMRA-labeled cyclophilin probe (), Texas Red–labeled IL-2 probe (). RFU, relative fluorescence units.

Reagents Application-Specific Kits and Reagents
Reagents
Application-Specific Kits and Reagents

Precision Melt Supermix

■■ Optimized formulation containing EvaGreen dye delivers robust PCR and high resolution melt (HRM) performance

■■ Sensitive and effective discrimination of all 4 SNP classes across a broad 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

A

0.20 0.15 0.10 0.05 0.00 –0.05 Difference RFU
0.20
0.15
0.10
0.05
0.00
–0.05
Difference RFU

78

79

80

81

82

Temperature, °C

83

B

0.02 0.00 –0.02 –0.04 –0.06 –0.08 Difference RFU Difference RFU
0.02
0.00
–0.02
–0.04
–0.06
–0.08
Difference RFU
Difference RFU

75

76

77

78

79

Temperature, °C

80

0.0

–0.1

–0.2

–0.3

–0.4

–0.5

–0.6

°C 80 0.0 –0.1 –0.2 –0.3 –0.4 –0.5 –0.6 74 76 78 80 Temperature, °C 82

74

76

78

80

Temperature, °C

82

84

Precision melt supermix delivers robust HRM for SNPs. Discrimination of class I and IV SNP genotypes are shown in panels A and B, respectively. Class I (A to G substitution) and class IV (A to T substitution) SNP genotypes from mouse genomic DNA were analyzed using precision melt supermix. Wild type (), heterozygote (), and homozygous mutant () are shown in the difference plots normalized to wild-type samples. HRM analysis was performed on a CFX384 real-time PCR detection system and genotypes were automatically assigned by Precision Melt Analysis software. Amplification was carried out for 35 cycles. Total run time including melt curve = 150 min. RFU, relative fluorescence units.

Accurate methylation detection with precision melt supermix. Mixtures of methylated and unmethylated human genomic DNA of varying ratios were analyzed using HRM on a CFX384 real-time PCR detection system. Increasing amounts of methylated DNA (, 0%; , 2%; , 5%; , 50%; , 75%; , 95%; and , 100%) were analyzed for methylation of the human RARB2 gene. The 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.

Reagents Application-Specific Kits and Reagents
Reagents
Application-Specific Kits and Reagents

EpiQ Chromatin Analysis Kit

■■ Novel technique generates quantitative chromatin structure information with strong correlation to gene expression levels

■■ Quantitative assessment of chromatin structure of target genes in cultured cells

■■ Kit discriminates open, actively transcribed chromatin regions from closed, transcriptionally silent regions

■■ In situ chromatin digestion, genomic DNA purification, and real-time PCR all combined in one workflow

■■ Short assay time — assessment of chromatin structure can be accomplished in less than 6 hr

■■ Small sample requirement — as little as 5 x 10 4 cells are required to perform analysis

EpiQ Chromatin SYBR ® Green Supermix

■■ Robust formulation delivers superior sensitivity and efficiency for qPCR from genomic DNA templates

■■

Protocol is optimized for difficult real-time qPCR reactions for high GC amplicons

■■ Supermix contains fluorescein and ROX and is compatible with all real-time PCR instruments except Applied Biosystems 7000, 7300, 7700, and 7900 models (additional ROX can be added by ordering the dye separately, catalog #172-5858)

Reagents B. GAPDH — Target Gene (constitutively expressed) 10 4 — No nuclease — With
Reagents
B. GAPDH — Target Gene (constitutively expressed)
10
4
— No nuclease
— With nuclease
10
3
10
2
ΔCq target = 8.08
10
1
RFU

A. HBB — Reference Gene (epigenetically silenced)

10 4 — No nuclease — With nuclease 10 3 10 2 ΔCq ref =
10
4
— No nuclease
— With nuclease
10
3
10
2
ΔCq ref = 0.58
10
1
RFU
Closed chromatin Open chromatin
Closed chromatin
Open chromatin

0

10

20

Cycles

30

40

0

10

20

Cycles

30

40

Chromatin consists of DNA spooled around complexes of histone protein molecules called nucleosomes ().

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.

Reagents Standard PCR Reagents
Reagents
Standard PCR Reagents

iTaq DNA Polymerase

■■ Antibody-mediated hot-start DNA polymerase for quick 3 min activation at 95°C

■■ Polymerase prevents nonspecific amplification and primer-dimers in both PCR and real-time PCR applications

dNTP Mix

■■ Formulated for consistency and higher efficiency in PCR and real-time PCR

■■

Robust dNTP solution withstands multiple rounds of freeze-thawing and temperature cycling

35 min iProof high-fidelity DNA polymerase 1 kb 2 hr Common high-fidelity polymerase 2 hr
35 min
iProof high-fidelity DNA polymerase
1
kb
2
hr
Common high-fidelity polymerase
2
hr
Taq polymerase
1.5 hr
8
kb
9.5 hr
5.5 hr
2 hr 20 min
15 kb
16.5 hr
(Failed amplification)
Template length

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.

iProof High-Fidelity DNA Polymerase

■■ A high-fidelity DNA polymerase with 52-fold more accuracy than Taq DNA polymerase

■■ Unique Pyrococcus-like proofreading enzyme is fused to a dsDNA binding protein, Sso7d

■■ Long and fast PCR applications — fragments up to 37 kb are amplified in less time (15–30 sec/kb) and with less enzyme (0.25–1 U/reaction)

■■ Convenient 2x supermix is available for iProof polymerase and buffer for GC-rich templates

BAC DNA (37 kb)

37 25 32 20 22 10 5
37
25
32
20
22
10
5

Human genomic DNA (28 kb)

22 28 15 7.9 3.8 2.9
22
28
15
7.9
3.8
2.9

– 48 kb

– 23 kb

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.

Reagents
Reagents
Real-Time qPCR Reagents Selection Guide
Real-Time qPCR Reagents Selection Guide

Reagents

   

SYBR ® Green / EvaGreen Supermixes

   

Probes Supermixes

 

One-Step Kits for RT- qPCR

SsoAdvanced

iTaq Universal SYBR ® Green Supermix

iQ SYBR ® Green Supermix

SsoFast

EpiQ Chromatin SYBR ® Green Supermix

SsoFast

iTaq

iQ

iQ

iScript One-Step RT-PCR Kit with SYBR ® Green

iScript

SYBR ®

EvaGreen ®

Probes

Universal

Supermix

Multiplex

One-Step

Green

Supermix

Supermix

Probes

Powermix

RT-PCR Kit

Real-Time PCR Instrument

Supermix*

 

Supermix

for Probes

Bio-Rad

                     

CFX96 , CFX96 Touch , CFX384 , CFX384 Touch , CFX Connect

✔ ✔

 

✔ ✔

 

✔ ✔

 

✔ ✔

   

✔ ✔

iQ , iQ 5, MyiQ , MyiQ 2

✔ ✔

 

✔ ✔

 

✔ ✔

 

✔ ✔

   

✔ ✔

MiniOpticon , DNA Engine 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

* SsoAdvanced universal SYBR ® Green supermix will be available soon.

ROX reference setting must be turned “off”

BSA must be added according to instrument specifications

Reagents Ordering Information
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 SsoAdvanced SYBR Green Supermix, 5 ml (5 x 1 ml vials), 500 x 20 μl reactions SsoAdvanced SYBR Green Supermix, 10 ml (10 x 1 ml vials), 1,000 x 20 μl reactions

172-5261

172-5262

172-5264

SsoAdvanced SYBR Green Supermix, 25 ml (5 x 5 ml vials), 2,500 x 20 μl reactions SsoAdvanced SYBR Green Supermix, 50 ml (10 x 5 ml vials), 5,000 x 20 μl reactions SsoFast Probes Supermix, 2 ml (2 x 1 ml vials), 200 x 20 μl reactions SsoFast Probes Supermix, 5 ml (5 x 1 ml vials), 500 x 20 μl reactions SsoFast Probes Supermix, 10 ml (10 x 1 ml vials), 1,000 x 20 μl reactions SsoFast Probes Supermix, 20 ml (20 ml bottle), 2,000 x 20 μl reactions

172-5265

172-5230

172-5231

172-5232

172-5233

iTaq Universal Supermixes

 

172-5120

iTaq Universal SYBR Green Supermix, 2 ml (2 x 1 ml vials), 200 x 20 μl reactions iTaq Universal SYBR Green Supermix, 5 ml (5 x 1 ml vials), 500 x 20 μl reactions iTaq Universal SYBR Green Supermix, 10 ml (10 x 1 ml vials), 1,000 x 20 μl reactions

172-5121

172-5122

172-5124

iTaq Universal SYBR Green Supermix, 25 ml (5 x 5 ml vials), 2,500 x 20 μl reactions iTaq Universal SYBR Green Supermix, 50 ml (10 x 5 ml vials), 5,000 x 20 μl reactions iTaq Universal Probes Supermix, 2 ml (2 x 1 ml vials), 200 x 20 μl reactions iTaq Universal Probes Supermix, 5 ml (5 x 1 ml vials), 500 x 20 μl reactions iTaq Universal Probes Supermix, 10 ml (10 x 1 ml vials), 1,000 x 20 μl reactions iTaq Universal Probes Supermix, 25 ml (5 x 5 ml vials), 2,500 x 20 μl reactions iTaq Universal Probes Supermix, 50 ml (10 x 5 ml vials), 5,000 x 20 μl reactions

172-5125

172-5130

172-5131

172-5132

172-5134

172-5135

iQ Supermixes

170-8880

iQ SYBR Green Supermix, 2.5 ml (2 x 1.25 ml vials), 100 x 50 μl reactions iQ SYBR Green Supermix, 12.5 ml (10 x 1.25 ml vials), 500 x 50 μl reactions iQ SYBR Green Supermix, 25 ml (20 x 1.25 ml vials), 1,000 x 50 μl reactions iQ SYBR Green Supermix, 50 ml (50 ml bottle), 2,000 x 50 μl reactions iQ SYBR Green Supermix, 25 ml (5 x 5 ml vials), 1,000 x 50 μl reactions iQ SYBR Green Supermix, 50 ml (10 x 5 ml vials), 2,000 x 50 μl reactions iQ Supermix, 2.5 ml (2 x 1.25 ml vials), 100 x 50 μl reactions iQ Supermix, 12.5 ml (10 x 1.25 ml vials), 500 x 50 μl reactions iQ Supermix, 25 ml (20 x 1.25 ml vials), 1,000 x 50 μl reactions iQ Multiplex Powermix, 1.25 ml (1 x 1.25 ml vial), 50 x 50 μl reactions iQ Multiplex Powermix, 5 ml (4 x 1.25 ml vials), 200 x 50 μl reactions

170-8882

170-8884

170-8885

170-8886

170-8887

170-8860

170-8862

170-8864

172-5848

172-5849

Reagents

Reagents Reagents Ordering Information Ordering Information   Catalog # Description $
Reagents Ordering Information
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 Precision Melt Supermix, 10 ml (10 x 1 ml vials), 1,000 x 20 μl reactions EpiQ Chromatin Analysis Kit, 50 preparations EpiQ Chromatin Analysis Kit, 100 preparations EpiQ Chromatin SYBR Green Supermix, 5 ml (5 x 1 ml vials), 500 x 20 μl reactions

172-5112

172-5400

172-5401

172-5404

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 iTaq DNA Polymerase, 5,000 U, 5 U/μl dNTP Mix, 200 μl iProof High-Fidelity DNA Polymerase, 20 U, 2 U/μl iProof High-Fidelity DNA Polymerase, 100 U, 2 U/μl iProof High-Fidelity DNA Polymerase, 500 U, 2 U/μl iProof HF Master Mix, 0.04 U/μl, 100 x 50 μl reactions iProof HF Master Mix, 0.04 U/μl, 500 x 50 μl reactions iProof GC Master Mix, 0.04 U/μl, 100 x 50 μl reactions

170-8875

170-8874

172-5300

172-5301

172-5302

172-5310

172-5311

172-5320

172-5321

iProof GC Master Mix, 0.04 U/μl, 500 x 50 μl reactions ROX Passive Reference Dye, 0.5 ml

172-5858

Additional Real-Time qPCR Supermixes*

 

172-5200

SsoFast EvaGreen Supermix, 2 ml (2 x 1 ml vials), 200 x 20 μl reactions SsoFast EvaGreen Supermix, 5 ml (5 x 1 ml vials), 500 x 20 μl reactions SsoFast EvaGreen Supermix, 10 ml (10 x 1 ml vials), 1,000 x 20 μl reactions SsoFast EvaGreen Supermix, 20 ml (20 ml bottle), 2,000 x 20 μl reactions SsoFast EvaGreen Supermix, 25 ml (5 x 5 ml vials), 2,500 x 20 μl reactions SsoFast EvaGreen Supermix, 50 ml (10 x 5 ml vials), 5,000 x 20 μl reactions

172-5201

172-5202

172-5203

172-5204

172-5205

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

PCR Plastic Consumables ■■ Precisely manufactured for optimal fit and cycling performance ■■ Produced in
PCR Plastic Consumables ■■ Precisely manufactured for optimal fit and cycling performance ■■ Produced in
PCR Plastic Consumables ■■ Precisely manufactured for optimal fit and cycling performance ■■ Produced in
PCR Plastic Consumables ■■ Precisely manufactured for optimal fit and cycling performance ■■ Produced in
PCR Plastic Consumables ■■ Precisely manufactured for optimal fit and cycling performance ■■ Produced in
PCR Plastic Consumables ■■ Precisely manufactured for optimal fit and cycling performance ■■ Produced in
PCR Plastic Consumables ■■ Precisely manufactured for optimal fit and cycling performance ■■ Produced in

PCR Plastic Consumables

■■ Precisely manufactured for optimal fit and cycling performance

■■ Produced in Class 10,000 or 100,000 cleanroom environment

■■ Certified to be free of DNase, RNase, and human genomic DNA

■■ Extremely uniform wells reduce well-to-well variability in real-time PCR

■■ Warp-free Hard-Shell ® plates are designed for optimum performance with automation

Plastics

PCR Plastic Consumables Instrument Compatibility 0.2 ml Tubes 384-Well Plates Individual High-Profile Strips
PCR Plastic Consumables
Instrument Compatibility
0.2 ml Tubes
384-Well Plates
Individual High-Profile
Strips High-Profile
Strips Low-Profile
Hard-Shell ® Standard
Hard-Shell 480
Catalog #
TBI-0201, TFI-0201, TWI-0201
TBS-xxxx, TBC-xxxx
TLS-xxxx
HSP-3xxx
HSR-48xx
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)

 

Applied Biosystems 384-well cyclers (9700, Veriti)

 

Eppendorf Mastercycler series

Real-Time PCR Instrument

 

Bio-Rad CFX96 , CFX96 Touch , CFX384 ,* CFX384 Touch ,* CFX Connect

 

Bio-Rad iCycler iQ ® , iQ 5, MyiQ , MyiQ 2

 

Bio-Rad Chromo4

Bio-Rad DNA Engine Opticon ® and 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

Roche LightCycler 480

Other Instruments

Applied Biosystems DNA sequencers (3100, 3700, 3730)

 

Idaho Technology LightScanner

 

Recommended

Compatible

PCR Plastic Consumables Instrument Compatibility
PCR Plastic Consumables
Instrument Compatibility
 

48- and 96-Well Plates

 

Hard-Shell ® Semi-Skirted High-Profile

Hard-Shell Skirted

Multiplate Unskirted High-Profile

Multiplate Unskirted

iQ Semi-Skirted High-Profile

Low-Profile

Low-Profile

Catalog #

HSS-xxxx

HSP-9xxx

MLP-xxxx

MLL-xxxx

223-9441

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 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 , CFX Connect

 

Bio-Rad iCycler iQ ® , iQ 5, MyiQ , MyiQ 2

Bio-Rad Chromo4

 

Bio-Rad DNA Engine Opticon ® , Opticon 2

 

Bio-Rad MiniOpticon *