The unique workflow of the NEBNext® Small RNA library prep kits addresses the challenge of minimization of adaptor-dimers while achieving production of high-yield, diverse multiplex libraries in a simple protocol.
The following reagents are supplied with this product:
NEB # | Component Name | Component # | Stored at (°C) | Amount | Concentration |
E7300S | -20 |
NEBNext 3´ Ligation Reaction Buffer | E7301AVIAL | -20 | 1 x 0.24 ml | ||
Murine RNase Inhibitor | E7308AVIAL | -20 | 1 x 0.024 ml | ||
NEBNext 5´ Ligation Reaction Buffer | E7304AVIAL | -20 | 1 x 0.024 ml | ||
NEBNext 5´ Ligation Enzyme Mix | E7305AVIAL | -20 | 1 x 0.06 ml | ||
LongAmp™ Taq 2X Master Mix | E7309AVIAL | -20 | 1 x 1.2 ml | ||
Quick-Load pBR322 DNA-MspI Digest | E7323AVIAL | -20 | 1 x 0.24 ml | ||
DNA Gel Elution Buffer | E7324AVIAL | -20 | 1 x 12 ml | ||
Linear Acrylamide | E7325AVIAL | -20 | 1 x 0.048 ml | ||
Nuclease-free Water | E7327AVIAL | -20 | 1 x 5 ml | ||
TE Buffer | E7326AVIAL | -20 | 1 x 0.48 ml | ||
NEBNext Index 1 Primer for Illumina | E7311AVIAL | -20 | 1 x 0.01 ml | 10 µM | |
NEBNext Index 2 Primer for Illumina | E7312AVIAL | -20 | 1 x 0.01 ml | 10 µM | |
NEBNext Index 3 Primer for Illumina | E7313AVIAL | -20 | 1 x 0.01 ml | 10 µM | |
NEBNext Index 4 Primer for Illumina | E7314AVIAL | -20 | 1 x 0.01 ml | 10 µM | |
NEBNext Index 5 Primer for Illumina | E7315AVIAL | -20 | 1 x 0.01 ml | 10 µM | |
NEBNext Index 6 Primer for Illumina | E7316AVIAL | -20 | 1 x 0.01 ml | 10 µM | |
NEBNext Index 7 Primer for Illumina | E7317AVIAL | -20 | 1 x 0.01 ml | 10 µM | |
NEBNext Index 8 Primer for Illumina | E7318AVIAL | -20 | 1 x 0.01 ml | 10 µM | |
NEBNext Index 9 Primer for Illumina | E7319AVIAL | -20 | 1 x 0.01 ml | 10 µM | |
NEBNext Index 10 Primer for Illumina | E7320AVIAL | -20 | 1 x 0.01 ml | 10 µM | |
NEBNext Index 11 Primer for Illumina | E7321AVIAL | -20 | 1 x 0.01 ml | 10 µM | |
NEBNext Index 12 Primer for Illumina | E7322AVIAL | -20 | 1 x 0.01 ml | 10 µM | |
Gel Loading Dye, Blue | E6138AVIAL | -20 | 1 x 0.2 ml | 6 X | |
NEBNext 3´ SR Adaptor for Illumina | E7332AVIAL | -20 | 1 x 0.024 ml | ||
NEBNext SR RT Primer for Illumina | E7333AVIAL | -20 | 1 x 0.024 ml | ||
NEBNext SR Primer for Illumina | E7310AVIAL | -20 | 1 x 0.06 ml | ||
NEBNext 5´ SR Adaptor for Illumina | E7328AVIAL | -20 | 1 x 1,350 pmol | ||
NEBNext First Strand Synthesis Reaction Buffer | E7334AVIAL | -20 | 1 x 0.192 ml | ||
ProtoScript II Reverse Transcriptase | E7355AVIAL | -20 | 1 x 0.024 ml | ||
NEBNext 3´ Ligation Enzyme Mix | E7288AVIAL | -20 | 1 x 0.072 ml |
E7300L | -20 |
NEBNext 3´ Ligation Reaction Buffer | E7301AAVIAL | -20 | 1 x 0.96 ml | ||
Murine RNase Inhibitor | E7308AAVIAL | -20 | 1 x 0.096 ml | ||
NEBNext 5´ Ligation Reaction Buffer | E7304AAVIAL | -20 | 1 x 0.096 ml | ||
NEBNext 5´ Ligation Enzyme Mix | E7305AAVIAL | -20 | 1 x 0.24 ml | ||
LongAmp™ Taq 2X Master Mix | E7309AAVIAL | -20 | 4 x 1.2 ml | ||
Quick-Load pBR322 DNA-MspI Digest | E7323AAVIAL | -20 | 1 x 0.96 ml | ||
DNA Gel Elution Buffer | E7324AAVIAL | -20 | 1 x 48 ml | ||
Linear Acrylamide | E7325AAVIAL | -20 | 1 x 0.192 ml | ||
Nuclease-free Water | E7327AAVIAL | -20 | 1 x 20 ml | ||
TE Buffer | E7326AAVIAL | -20 | 1 x 1.92 ml | ||
NEBNext Index 1 Primer for Illumina | E7311AAVIAL | -20 | 1 x 0.04 ml | 10 µM | |
NEBNext Index 2 Primer for Illumina | E7312AAVIAL | -20 | 1 x 0.04 ml | 10 µM | |
NEBNext Index 3 Primer for Illumina | E7313AAVIAL | -20 | 1 x 0.04 ml | 10 µM | |
NEBNext Index 4 Primer for Illumina | E7314AAVIAL | -20 | 1 x 0.04 ml | 10 µM | |
NEBNext Index 5 Primer for Illumina | E7315AAVIAL | -20 | 1 x 0.04 ml | 10 µM | |
NEBNext Index 6 Primer for Illumina | E7316AAVIAL | -20 | 1 x 0.04 ml | 10 µM | |
NEBNext Index 7 Primer for Illumina | E7317AAVIAL | -20 | 1 x 0.04 ml | 10 µM | |
NEBNext Index 8 Primer for Illumina | E7318AAVIAL | -20 | 1 x 0.04 ml | 10 µM | |
NEBNext Index 9 Primer for Illumina | E7319AAVIAL | -20 | 1 x 0.04 ml | 10 µM | |
NEBNext Index 10 Primer for Illumina | E7320AAVIAL | -20 | 1 x 0.04 ml | 10 µM | |
NEBNext Index 11 Primer for Illumina | E7321AAVIAL | -20 | 1 x 0.04 ml | 10 µM | |
NEBNext Index 12 Primer for Illumina | E7322AAVIAL | -20 | 1 x 0.04 ml | 10 µM | |
Gel Loading Dye, Blue | E6138AAVIAL | -20 | 1 x 1 ml | 6 X | |
NEBNext 3´ SR Adaptor for Illumina | E7332AAVIAL | -20 | 1 x 0.096 ml | ||
NEBNext SR RT Primer for Illumina | E7333AAVIAL | -20 | 1 x 0.096 ml | ||
NEBNext SR Primer for Illumina | E7310AAVIAL | -20 | 1 x 0.24 ml | ||
NEBNext 5´ SR Adaptor for Illumina | E7328AVIAL | -20 | 1 x 1,350 pmol | ||
NEBNext First Strand Synthesis Reaction Buffer | E7334AAVIAL | -20 | 1 x 0.768 ml | ||
ProtoScript II Reverse Transcriptase | E7355AAVIAL | -20 | 1 x 0.096 ml | ||
NEBNext 3´ Ligation Enzyme Mix | E7288AAVIAL | -20 | 1 x 0.288 ml |
3 M Sodium Acetate, pH 5.5
100% Ethanol
80% Ethanol
Corning®, Costar®, Spin-X® Centrifuge Tube Filters (Cellulose Acetate Filters) (Sigma Aldrich # CLS8162)
Magnetic rack or plate (e.g., NEBNext® Magnetic Separation Rack (NEB #S1515S), Alpaqua ® 96S Super Magnet Plate (#A001322), or equivalent)
Size Selection Materials
for gel size selection:
6% Novex® TBE PAGE gel, 1.0 mM
10-well (Life Technologies, Inc. #EC6265BOX)
SYBR® Gold Nucleic Acid Gel Stain (Life Technologies, Inc. #S-11494)
RNase-free Disposable Pellet Pestles® (Kimble Kontes Asset Management, Inc. #749521-1590)
Dry Ice/Methanol Bath or –80°C freezer
for bead selection:
Agencourt® AMPure® XP Beads (Beckman Coulter, Inc. #A63881)
for Pippin Prep™ selection:
3% Agarose Dye Free Gel (Sage Science #CDF 3010)
Monarch® PCR & DNA Cleanup Kit (5 μg) (NEB# T1030)
Bioanalyzer® (Agilent® Technologies, Inc.)
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NEW ENGLAND BIOLABS®, LongAmp®, NEBNext® and QUICK-LOAD® are registered trademarks of New England Biolabs, Inc.
AGENCOURT® and AMPure® are registered trademarks of Beckman Coulter, Inc.
Agilent® and Bioanalyzer® are registered trademarks of Agilent Technologies, Inc.
Illumina® is a registered trademark of Illumina, Inc.
CORNING®, COSTAR® and SPIN-X® are registered trademarks of Corning, Inc.
NOVEX®, FIRSTCHOICE® and SYBR® are registered trademarks of Life Technologies, Inc.
FLASHPAGE™ is a registered trademark of Life Technologies, Inc.
Pellet Pestle® is a registered trademark of Kimble Kontes Asset Management, Inc.
Qiagen® and QIAQUICK® are registered trademarks of The Qiagen Group.