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NEBNext® Multiplex Small RNA Library Prep Set for Illumina® (Set 2)

Catalog # Concentration Size List Price Quantity Your Price
E7580L 96 reactions $7,765.00
$6,988.50
E7580S 24 reactions $2,285.00
$2,056.50
Catalog # Size List Price Your Price
E7580L 96 reactions $7,765.00
$6,988.50
E7580S 24 reactions $2,285.00
$2,056.50
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

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. 

  • Minimized adaptor-dimer contamination
  • High yields
  • Input RNA can be Total RNA
  • Suitable for methylated small RNAs (e.g. piRNAs) as well as unmethylated small RNAs
Featured Videos
View Video Library
  • Monarch_MiniprepProtocol_VideoThumb
    A Breakthrough Method of Small RNA Sample Preparation for Next Gen Sequencing
  • Monarch_MiniprepProtocol_VideoThumb
    Tips for Optimizing RNA Inputs in NGS Library Construction
  • Monarch_MiniprepProtocol_VideoThumb
    12 Quick Tips for NGS Library Preparation
  • Monarch_MiniprepProtocol_VideoThumb
    NEBNEXT® for Core Lab Facilities


The novel NEBNext Small RNA workflow has been optimized to minimize adaptor-dimers while producing high-yield, high-diversity libraries. The NEBNext Small RNA Library Prep Set for Illumina® (Set 2) includes adaptors and multiplex primers with 12 indices, to enable multiplexing. Additional index primers for multiplexing are included in kits NEB #E7300 and NEB #E7560.

Each kit component must pass rigorous quality control standards, and each set of reagents is functionally validated together by construction and sequencing of indexed small RNA libraries on the Illumina sequencing platform.

Small RNA Library Preparation Workflow for Illumina



GenomeWeb article reviews NEBNext Small RNA Kits.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  NEBNext 3´ Ligation Reaction Buffer E7301AVIAL -20 1 x 0.24 ml
  NEBNext 3´ SR Adaptor for Illumina E7332AVIAL -20 1 x 0.024 ml
  NEBNext 5´ SR Adaptor for Illumina E7328AVIAL -20 1 x 1,350 pmol
  NEBNext 5´ Ligation Reaction Buffer E7304AVIAL -20 1 x 0.024 ml
  NEBNext 5´ Ligation Enzyme Mix E7305AVIAL -20 1 x 0.06 ml
  NEBNext SR RT Primer for Illumina E7333AVIAL -20 1 x 0.024 ml
  NEBNext First Strand Synthesis Reaction Buffer E7334AVIAL -20 1 x 0.192 ml
  ProtoScript II Reverse Transcriptase E7355AVIAL -20 1 x 0.024 ml
  Murine RNase Inhibitor E7308AVIAL -20 1 x 0.024 ml
  LongAmp™ Taq 2X Master Mix E7309AVIAL -20 1 x 1.2 ml
  NEBNext SR Primer for Illumina E7310AVIAL -20 1 x 0.06 ml
  Gel Loading Dye, Blue E6138AVIAL -20 1 x 0.2 ml 6 X
  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
  TE Buffer E7326AVIAL -20 1 x 0.48 ml
  Nuclease-free Water E7327AVIAL -20 1 x 5 ml
  NEBNext Index 13 Primer for Illumina E7581AVIAL -20 1 x 0.01 ml 10 µM
  NEBNext Index 14 Primer for Illumina E7582AVIAL -20 1 x 0.01 ml 10 µM
  NEBNext Index 15 Primer for Illumina E7583AVIAL -20 1 x 0.01 ml 10 µM
  NEBNext Index 16 Primer for Illumina E7584AVIAL -20 1 x 0.01 ml 10 µM
  NEBNext Index 17 Primer for Illumina E7585AVIAL -20 1 x 0.01 ml 10 µM
  NEBNext Index 18 Primer for Illumina E7586AVIAL -20 1 x 0.01 ml 10 µM
  NEBNext Index 19 Primer for Illumina E7587AVIAL -20 1 x 0.01 ml 10 µM
  NEBNext Index 20 Primer for Illumina E7588AVIAL -20 1 x 0.01 ml 10 µM
  NEBNext Index 21 Primer for Illumina E7589AVIAL -20 1 x 0.01 ml 10 µM
  NEBNext Index 22 Primer for Illumina E7590AVIAL -20 1 x 0.01 ml 10 µM
  NEBNext Index 23 Primer for Illumina E7591AVIAL -20 1 x 0.01 ml 10 µM
  NEBNext Index 24 Primer for Illumina E7592AVIAL -20 1 x 0.01 ml 10 µM
  NEBNext 3´ Ligation Enzyme Mix E7288AVIAL -20 1 x 0.072 ml
  NEBNext 3´ Ligation Reaction Buffer E7301AAVIAL -20 1 x 0.96 ml
  NEBNext 3´ SR Adaptor for Illumina E7332AAVIAL -20 1 x 0.096 ml
  NEBNext 5´ SR Adaptor for Illumina E7328AVIAL -20 1 x 1,350 pmol
  NEBNext 5´ Ligation Reaction Buffer E7304AAVIAL -20 1 x 0.096 ml
  NEBNext 5´ Ligation Enzyme Mix E7305AAVIAL -20 1 x 0.24 ml
  NEBNext SR RT Primer for Illumina E7333AAVIAL -20 1 x 0.096 ml
  NEBNext First Strand Synthesis Reaction Buffer E7334AAVIAL -20 1 x 0.768 ml
  ProtoScript II Reverse Transcriptase E7355AAVIAL -20 1 x 0.096 ml
  Murine RNase Inhibitor E7308AAVIAL -20 1 x 0.096 ml
  LongAmp™ Taq 2X Master Mix E7309AAVIAL -20 4 x 1.2 ml
  NEBNext SR Primer for Illumina E7310AAVIAL -20 1 x 0.24 ml
  Gel Loading Dye, Blue E6138AAVIAL -20 1 x 1 ml 6 X
  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
  TE Buffer E7326AAVIAL -20 1 x 1.92 ml
  Nuclease-free Water E7327AAVIAL -20 1 x 20 ml
  NEBNext Index 13 Primer for Illumina E7581AAVIAL -20 1 x 0.04 ml 10 µM
  NEBNext Index 14 Primer for Illumina E7582AAVIAL -20 1 x 0.04 ml 10 µM
  NEBNext Index 15 Primer for Illumina E7583AAVIAL -20 1 x 0.04 ml 10 µM
  NEBNext Index 16 Primer for Illumina E7584AAVIAL -20 1 x 0.04 ml 10 µM
  NEBNext Index 17 Primer for Illumina E7585AAVIAL -20 1 x 0.04 ml 10 µM
  NEBNext Index 18 Primer for Illumina E7586AAVIAL -20 1 x 0.04 ml 10 µM
  NEBNext Index 19 Primer for Illumina E7587AAVIAL -20 1 x 0.04 ml 10 µM
  NEBNext Index 20 Primer for Illumina E7588AAVIAL -20 1 x 0.04 ml 10 µM
  NEBNext Index 21 Primer for Illumina E7589AAVIAL -20 1 x 0.04 ml 10 µM
  NEBNext Index 22 Primer for Illumina E7590AAVIAL -20 1 x 0.04 ml 10 µM
  NEBNext Index 23 Primer for Illumina E7591AAVIAL -20 1 x 0.04 ml 10 µM
  NEBNext Index 24 Primer for Illumina E7592AAVIAL -20 1 x 0.04 ml 10 µM
  NEBNext 3´ Ligation Enzyme Mix E7288AAVIAL -20 1 x 0.288 ml

Properties & Usage

Materials Required but not Supplied

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.)

Citations
Additional Citations
  • Macchiaroli, N. et al. (2015) microRNA profiling in the zoonotic parasite Echinococcus canadensis using a high-throughput approach. Parasit Vectors 8, 83.PubMedID: 25656283
  • Tsai, H.C. et al (2015) Transcriptional Analysis of Deinococcus radiodurans Reveals Novel Small RNAs That Are Differentially Expressed under Ionizing Radiation. Appl Environ Microbiol 81, 1754-1764.PubMedID: 25548054
Quality Control Assay
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
Specifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
Certificate of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Safety Data Sheets
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This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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