Beruflich Dokumente
Kultur Dokumente
Table of Contents:
Applications...................................................................................................3
NEBNext Adaptor for Illumina........................................................................5
USER Enzyme.......................................................................................................6
NEBNext Universal PCR Primer for Illumina..........................................................7
NEBNext Index 112 Primers for Illumina..................................................8-9
Revision History..........................................................................................10
Note: Concentrations of Universal PCR Primer and the Index Primers have
changed from 25 M to 10 M. Please refer to the most recent protocol
revisions for use.
Applications:
The NEBNext Multiplex Oligos for Illumina (Index Primers Set 1) contains
adaptors and primers that are ideally suited for multiplex sample
preparation for next-generation sequencing on the Illumina platform
(Illumina, Inc.). Each of these components must pass rigorous quality control
standards and is lot controlled, both individually and as a set of reagents.
Lot Control: The lots provided in the NEBNext Multiplex Oligos for Illumina
(Index Primers Set 1) are managed separately and are qualified by
additional functional validation. Individual reagents undergo standard
enzyme activity and quality control assays, and also meet stringent criteria
in the additional quality controls listed on each individual component page.
Functionally Validated: Each set of reagents is functionally validated together
through construction and sequencing of genomic DNA libraries as well as
human mRNA libraries and ChIP seq libraries on the Illumina platform.
For larger volume requirements, customized and bulk packaging is available by
purchasing through the OEM/Bulks department at NEB. Please contact OEM@
neb.com for further information.
Figure 1: Workflow.
A
A
End repaired,
dA-tailed DNA fragments
T
A
USER treatment
USER
PCR amplification
#E7420, NEBNext Ultra Directional RNA Library Prep Kit for Illumina
#E6040, NEBNext DNA Library Prep Master Mix Set for Illumina
#E6110, NEBNext mRNA Library Prep Master Mix Set for Illumina
#E6240, NEBNext ChIP-Seq Library Prep Master Mix Set for Illumina
0.24 ml
0.96 ml
Concentration: 15 M
5-/5Phos/GAT CGG AAG AGC ACA CGT CTG AAC TCC AGT C/ideoxyU/A CAC TCT TTC CCT ACA CGA
CGC TCT TCC GAT C*T-3
Store at 20C
U.S. PATENTS: 8,420,319, 8,288,097. PENDING U.S. AND FOREIGN APPLICATIONS: US 20120244525, US
20120238738, WO 2012012037.
USER Enzyme
#E7338A: 0.072 ml
#E7338AA: 0.288 ml
Store at 20C
Supplied in: 50 mM KCl, 5 mM NaCl, 10 mM Tris-HCl (pH 7.4 @ 25C),
0.1 mM EDTA, 1 mM DTT, 175 g/ml BSA and 50% Glycerol
Concentration: 10 M
5-AAT GAT ACG GCG ACC ACC GAG ATC TAC ACT CTT TCC CTA CAC GAC GCT CTT CCG ATC*T-3
Store at 20C
#E7311A:
ml
#E7311AA:
ml
#E7312A:
ml
#E7312AA:
ml
#E7313A:
ml
#E7313AA:
ml
#E7314A:
ml
#E7314AA:
ml
#E7315A:
ml
#E7315AA:
ml
#E7316A:
ml
#E7316AA:
ml
#E7317A:
ml
#E7317AA:
ml
#E7318A:
ml
#E7318AA:
ml
#E7319A:
ml
#E7319AA:
ml
#E7320A:
ml
#E7320AA:
ml
#E7321A:
ml
#E7321AA:
ml
#E7322A:
ml
#E7322AA:
ml
0.010
0.040
0.010
0.040
0.010
0.040
0.010
0.040
0.010
0.040
0.010
0.040
0.010
0.040
0.010
0.040
0.010
0.040
0.010
0.040
0.010
0.040
0.010
0.040
NEBNext
Index 1
Primer for
Illumina
NEBNext
Index 2
Primer for
Illumina
NEBNext
Index 3
Primer for
Illumina
NEBNext
Index 4
Primer for
Illumina
NEBNext
Index 5
Primer for
Illumina
NEBNext
Index 6
Primer for
Illumina
NEBNext
Index 7
Primer for
Illumina
NEBNext
Index 8
Primer for
Illumina
NEBNext
Index 9
Primer for
Illumina
NEBNext
Index 10
Primer for
Illumina
NEBNext
Index 11
Primer for
Illumina
NEBNext
Index 12
Primer for
Illumina
5CAAGCAGAAGACGGCATACGAGATCGTGATGTGACTGGAGTTCAGACGTGTGCTCTTCCGATC-s-T-3
5CAAGCAGAAGACGGCATACGAGATACATCGGTGACTGGAGTTCAGACGT
- GTGCTCTTCCGATC-s-T-3
5CAAGCAGAAGACGGCATACGAGATGCCTAAGTGACTGGAGTTCAGACG
T- GTGCTCTTCCGATC-s-T-3
5CAAGCAGAAGACGGCATACGAGATTGGTCAGTGACTGGAGTTCAGACGTGTGCTCTTCCGATC-s-T-3
5CAAGCAGAAGACGGCATACGAGATCACTGTGTGACTGGAGTTCAGACGTGTGCTCTTCCGATC-s-T-3
5CAAGCAGAAGACGGCATACGAGATATTGGCGTGACTGGAGTTCAGA
CGT- GTGCTCTTCCGATC-s-T-3
5CAAGCAGAAGACGGCATACGAGATGATCTGGTGACTGGAGTTCAGA
CGT- GTGCTCTTCCGATC-s-T-3
5CAAGCAGAAGACGGCATACGAGATTCAAGTGTGACTGGAGTTCAGA
CGT- GTGCTCTTCCGATC-s-T-3
5CAAGCAGAAGACGGCATACGAGA
TCTGATCGTGACTGGAGTTCAGA
5CAAGCAGAAGACGGCATACGAGATAAGCTAGTGACTGGAGTTCAGACGT
- GTGCTCTTCCGATC-s-T-3
5CAAGCAGAAGACGGCATACGAGATGTAGCCGTGACTGGAGTTCAGA
CGT- GTGCTCTTCCGATC-s-T-3
5CAAGCAGAAGACGGCATACGAGATTACAAGGTGACTGGAGTTCAGACGT
- GTGCTCTTCCGATC-s-T-3
Expe
cted
Inde
x
Prim
er
ATCACG
CGATGT
TTAGGC
TGACCA
ACAGT
G
GCCAAT
CAGATC
ACTTGA
GATCA
G
TAGCTT
GGCTA
C
CTTGTA
Concentration: 10 M
Revision History:
10
Revision #
Description
3.0
11
DNA CLONING
DNA AMPLIFICATION & PCR
EPIGENETICS
RNA ANALYSIS
LIBRARY PREP FOR NEXT GEN SEQUENCING
PROTEIN EXPRESSION & ANALYSIS
CELLULAR ANALYSIS
USA
Version 3.0
1/15