NEBNext® Small RNA Library Prep Set for SOLiD™

  • This product was discontinued on 01/19/2016
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Description

The NEBNext Small RNA Library Prep Set for SOLiD™ contains adaptors, primers, enzymes and buffers that are ideal to convert small RNA into cDNA libraries for next-generation sequencing on the SOLiD platform. NEBNext Small RNA Library Prep Set for Solid is multiplex compatible. Each of these components must pass rigorous quality control standards and are lot controlled, both individually and as a set of reagents.

Functional Validation
Each set of reagents is functionally validated together through construction and sequencing of an RNA library on the SOLiD™ 4.

Recent article in GenomeWeb reviews NEBNext Small RNA Kits.

Lot Control

The lots provided are managed separately and 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

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C)Concentration
ProtoScript® II Reverse Transcriptase-20200,000 units/ml
T4 DNA Ligase Reaction Buffer-2010X
NEBNext 3' Ligation Reaction Buffer-202X
NEBNext 3´ Ligation Enzyme Mix-20
NEBNext 3' SR Adaptor 3-20
NEBNext 5' SR Adaptor 3-20
NEBNext 5' Ligation Reaction Buffer-2010X
NEBNext 5' Ligation Enzyme Mix-20
NEBNext SR RT Primer 3-20
NEBNext First Strand Synthesis Reaction Buffer-205X
Murine RNase Inhibitor-20
OneTaq Hot Start 2X Master Mix with Standard Buffer-202X
NEBNext Index 1 Primer for SOLiD-20
NEBNext SR Primer R3-20
Gel Loading Dye, Blue256X
Quick-Load pBR322 DNA-MspI Digest-20100 μg/ml
DNA Gel Elution Buffer41X
TE Buffer-20
Nuclease-free Water-20
Linear Acrylamide-2010 mg/ml

Properties and Usage

Materials Required but not Supplied


3 M Sodium Acetate, pH 5.2
100% Ethanol
80% Ethanol
SuperScript® II Reverse Transcriptase (Invitrogen #18064-014)
5X First Strand Buffer (Supplied with SuperScript® II Reverse Transcriptase)
0.1 M DTT (Supplied with SuperScript® II Reverse Transcriptase)
Costar Spin-X centrifuge tube Filters (Cellulose Acetate Filters) (Sigma #CLS8162)
RNase-free Disposable Pellet Pestles (Kontes #74951-1590)
6% Novex TBE PAGE gel, 1.0 mm, 10 well (Invitrogen #EC6265BOX)
SYBR® Gold Nucleic Acid Gel Stain (Invitrogen #S-11494)

Storage Temperature

-20°C

FAQs

  1. Is Index 1 in the NEBNext Small RNA Library Prep Set for SOLiD kit (E6160) compatible with the NEBNext Small RNA Multiplex (1-16) Prep Set for SOLiD kit (E7450)?
  2. If my input is <5ug and therefore use 0.5 ul of  3' SR Adapter 3, SR RT Primer 3 and 5' SR Adapter 3, instead of 1.0 ul, my total volumes for each reaction will be short by 0.5 ul. Should I replace that lost volume with water?
  3. Why do I add the SR RT Primer 3 before the 5’ ligation instead of right before the reverse transcription step?
  4. If I forget to add the 3’ SR RT Primer 3 before the 5’ ligation can I add it before the reverse transcription step and move forward?
  5. Can samples be stored longer than 72 hours at -20°C?
  6. How many cycles of PCR should I do?
  7. If I accidentally move all 30 ul of the 5’ ligation product forward to the reverse transcription mixture will the reaction still work?

Protocols

  1. Library Preparation Using NEBNext® Small RNA Library Prep Set for SOLiD™ (E6160)

Manuals

The Product Manual includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name these document files: manual[Catalog Number].

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Safety Data Sheet

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.