Product Class:

NEBNext® Multiplex Small RNA Library Prep Set for Illumina® (Set 1)
Item# Description List Price Web Price Qty
E7300L NEBNext Multiplex Small RNA Library Prep Set for Illumina (1-12) - 96 rxns $7,239.00
$6,515.10
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E7300S NEBNext Multiplex Small RNA Library Prep Set for Illumina (1-12) - 24 rxns $2,129.00
$1,916.10
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*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

Note that additional index primers for multiplexing are provided with kit NEB #E7580 and NEB #E7560.

Product Introduction

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

Product Information

Description







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 1) includes adaptors and multiplex primers with 12 indices, to enable multiplexing. Additional index primers for multiplexing are included in kits NEB #E7580 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.

For larger volume requirements, customized and bulk packaging is available by purchasing through the OEM/Bulks department at NEB. Please contact [email protected] for further information.

Small RNA Library Preparation Workflow for Illumina

GenomeWeb article reviews NEBNext Small RNA Kits.

Each set of reagents is functionally validated together through construction and sequencing of a transcriptome library and sequenced on a

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
Gel Loading Dye, Blue 25 6 X
NEBNext 3´ Ligation Enzyme Mix -20
NEBNext 3´ Ligation Reaction Buffer -20 2 X
NEBNext 5´ Ligation Reaction Buffer -20
NEBNext 5´ Ligation Enzyme Mix -20
Murine RNase Inhibitor
LongAmp™ Taq 2X Master Mix -20 2 X
NEBNext SR Primer for Illumina -20
NEBNext Index 1 Primer for Illumina -20
NEBNext Index 2 Primer for Illumina -20
NEBNext Index 3 Primer for Illumina -20
NEBNext Index 4 Primer for Illumina -20
NEBNext Index 5 Primer for Illumina -20
NEBNext Index 6 Primer for Illumina -20
NEBNext Index 7 Primer for Illumina -20
NEBNext Index 8 Primer for Illumina -20
NEBNext Index 9 Primer for Illumina -20
NEBNext Index 10 Primer for Illumina -20
NEBNext Index 11 Primer for Illumina -20
NEBNext Index 12 Primer for Illumina -20
Quick-Load pBR322 DNA-MspI Digest -20 50 µg/ml
DNA Gel Elution Buffer -20 1 X
Linear Acrylamide -20 10 mg/ml
TE Buffer -20
Nuclease-free Water -20
NEBNext 5´ SR Adaptor for Illumina -20
NEBNext 3´ SR Adaptor for Illumina -20
NEBNext SR RT Primer for Illumina -20
NEBNext First Strand Synthesis Reaction Buffer -20 5 X
ProtoScript II Reverse Transcriptase -20 200000 units/ml
Applications:
RNA For Illumina,
RNA-seq,
Illumina Library Preparation

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.)

Storage Temperature

-20°C

Product Notes

  1. Kit Optimization

    The components of this kit were collectively optimized with FirstChoice®Human Brain Reference RNA (Life Technologies, Inc. #AM7962).

    High quality total RNA should be used as starting material. An RNA IntegrityNumber (RIN) > 7 is recommended.

    Assess the quality and quantity of your sample using the Agilent 2100Bioanalyzer using an Agilent RNA 6000 Nano Chip.

Protocols, Manuals & Usage

Protocols

  1. Protocol for use with NEBNext Small RNA Library Prep Set for Illumina (E7300, E7580, E7560, E7330)
  2. 6A. Quality Control Check and Size Selection using 6% Polyacrylamide Gel NEBNext Small RNA Library Prep Set for Illumina (E7300, E7580, E7560, E7330)
  3. 6B. Quality Control Check and Size Selection using Pippin Prep NEBNext Small RNA Library Prep Set for Illumina (E7300, E7580, E7560, E7330)
  4. 6C. Quality Control Check and Size Selection using AMPure XP Beads NEBNext Small RNA Library Prep Set for Illumina (E7300, E7580, E7560, E7330)

Manuals

The Product Manual includes details for how to use the product, as well as details of its formulation and quality controls.

Application Notes

Faqs & Troubleshooting

FAQs

  1. Is it possible to prevent abundant, unwanted RNAs to be included in the Small RNA library?
  2. Can I use Total RNA to make small RNA libraries or do I have to isolate or enrich the sample for small RNA?
  3. Why does the RT primer hybridization occur before 5’adaptor ligation?
  4. Do I have to hybridize the RT primer again after 5’ ligation?
  5. How are the barcodes introduced in the Multiplex libraries?
  6. During size selection on 6% PAGE gel, which bands should I cut out of the gel?
  7. Are libraries prepared by this method compatible with paired-end flowcells for cluster generation?
  8. Can I use the small RNA sample preparation kit for Directional-RNA sequencing?
  9. What is the sequence of the final PCR product?
  10. Can I use enriched small RNA instead of total RNA for the NEBNext Small RNA Library Prep for Illumina?
  11. What should I do if my Small RNA may not have 5´ Phosphate and 3´ OH groups?
  12. How should my NEBNext Small RNA library be trimmed?
  13. Are the Monarch RNA Cleanup Kits compatible with NEBNext reagents for RNA library prep?
  14. What should I do if the sample sheet from the NEB.com website is not compatible with the software version I am using?

Troubleshooting

Quality, Safety & Legal

Quality Assurance Statement

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 DataSheets

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

Legal and Disclaimers

This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.

For more information about commercial rights, please contact NEB's Global Business Development team at [email protected].

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.

Trademarks


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.