Product Class: Kit

NEBNext® Multiplex Oligos for Illumina® (Index Primers Set 3)

Product Introduction

NEBNext® Multiplex Oligos provide adaptors and primers to enable high yield multiplex Illumina library production. The unique hairpin loop structure of the NEBNext Adaptor minimizes adaptor-dimer formation, and NEBNext index PCR primers enable index incorporation during library amplification.

For additional information on available indices and formats for multiplexing, please refer to the NEBNext Multiplex Oligos Selection Chart.

  • High efficiency adaptor ligation
  • Minimized adaptor-dimer formation
  • Includes index primers for library multiplexing

Note: NEBNext® Multiplex Oligos for Illumina® (Index Primer Set 1) (NEB #7335) includes 12 indices. 

 
Catalog # Size Concentration
E7710S 24.0 reactions (12.0 indices)
E7710L 96.0 reactions (12.0 indices)

Product Information

Description

Incorporating a novel hairpin loop structure, the NEBNext Adaptor ligates with increased efficiency to end-repaired, dA-tailed DNA. The loop contains a U, which is removed by treatment with USER® Enzyme (a mix of UDG and Endo VIII), to open up the loop and make it available as a substrate for PCR. During PCR, barcodes can be incorporated by use of the NEBNext index primers, thereby enabling multiplexing.

NEBNext Oligos can be used with NEBNext products, and with other standard Illumina-compatible library preparation protocols.

Additional single- or dual-barcode primer options are available, in 12- and 96- index formats (NEB #E7335, NEB #E7500, NEB #E7730, NEB #E6609, NEB #E7600). A methylated version of the NEBNext Adaptor is also available for use with bisulfite sequencing protocols (NEB #E7535).


Figure 1: Workflow demonstrating the use of NEBNext Multiplex Oligos for Illumina (Index Primers Set 3)

Figure 1: Workflow


This product is related to the following categories:
NEBNext® Multiplex Oligos (Adaptors & Primers) Products,
Next Generation Sequencing Library Preparation Products,
This product can be used in the following applications:
Applications of USER® and Thermolabile USER II Enzymes

Kit Components

Kit Components

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • E7710S     -20    
      NEBNext Index 26 Primer for Illumina E7713AVIAL -20 1 x 0.01 ml 10 µM
      NEBNext Index 28 Primer for Illumina E7714AVIAL -20 1 x 0.01 ml 10 µM
      NEBNext Index 29 Primer for Illumina E7715AVIAL -20 1 x 0.01 ml 10 µM
      NEBNext Index 31 Primer for Illumina E7717AVIAL -20 1 x 0.01 ml 10 µM
      NEBNext Index 32 Primer for Illumina E7718AVIAL -20 1 x 0.01 ml 10 µM
      NEBNext Index 33 Primer for Illumina E7719AVIAL -20 1 x 0.01 ml 10 µM
      NEBNext Index 34 Primer for Illumina E7720AVIAL -20 1 x 0.01 ml 10 µM
      NEBNext Index 35 Primer for Illumina E7721AVIAL -20 1 x 0.01 ml 10 µM
      NEBNext Index 36 Primer for Illumina E7722AVIAL -20 1 x 0.01 ml 10 µM
      NEBNext Universal PCR Primer for Illumina E6861AVIAL -20 1 x 0.12 ml 10 µM
      NEBNext Adaptor for Illumina E7337AVIAL -20 1 x 0.24 ml Not Applicable
      USER® Enzyme E7338AVIAL -20 1 x 0.072 ml Not Applicable
      NEBNext Index 42 Primer for Illumina E7736AVIAL -20 1 x 0.01 ml 10 µM
      NEBNext Index 24 Primer for Illumina E7592AVIAL -20 1 x 0.01 ml 10 µM
      NEBNext Index 17 Primer for Illumina E7585AVIAL -20 1 x 0.01 ml 10 µM
  • E7710L     -20    
      NEBNext Adaptor for Illumina E7337AAVIAL -20 1 x 0.96 ml Not Applicable
      USER® Enzyme E7338AAVIAL -20 1 x 0.288 ml Not Applicable
      NEBNext Universal PCR Primer for Illumina E6861AAVIAL -20 1 x 0.48 ml 10 µM
      NEBNext Index 17 Primer for Illumina E7585AAVIAL -20 1 x 0.04 ml 10 µM
      NEBNext Index 24 Primer for Illumina E7592AAVIAL -20 1 x 0.04 ml 10 µM
      NEBNext Index 26 Primer for Illumina E7713AAVIAL -20 1 x 0.04 ml 10 µM
      NEBNext Index 28 Primer for Illumina E7714AAVIAL -20 1 x 0.04 ml 10 µM
      NEBNext Index 29 Primer for Illumina E7715AAVIAL -20 1 x 0.04 ml 10 µM
      NEBNext Index 31 Primer for Illumina E7717AAVIAL -20 1 x 0.04 ml 10 µM
      NEBNext Index 32 Primer for Illumina E7718AAVIAL -20 1 x 0.04 ml 10 µM
      NEBNext Index 33 Primer for Illumina E7719AAVIAL -20 1 x 0.04 ml 10 µM
      NEBNext Index 34 Primer for Illumina E7720AAVIAL -20 1 x 0.04 ml 10 µM
      NEBNext Index 35 Primer for Illumina E7721AAVIAL -20 1 x 0.04 ml 10 µM
      NEBNext Index 36 Primer for Illumina E7722AAVIAL -20 1 x 0.04 ml 10 µM
      NEBNext Index 42 Primer for Illumina E7736AAVIAL -20 1 x 0.04 ml 10 µM

Properties & Usage

Materials Required but not Supplied

For the list of additional materials required, please check the manual for your NEBNext Library Prep Kit.

Protocols, Manuals & Usage

Manuals

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

Usage & Guidelines

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. Do you have a Sample Sheet with the index sequences for NEBNextMultiplex Oligos for Illumina (Set 3, #E7710) that can be used in theIllumina Experiment Manager (IEM)?
  2. Are NEBNext adaptors and primers for Illumina validated in Next Generation Sequencing Workflows?
  3. Are NEBNext adaptor and primers for Illumina compatible with NEBNext reagents for Illumina library preparation?
  4. Is USER enzyme required for library preparation using NEBNext adaptor and primers for Illumina?
  5. How much DNA can I ligate to the NEBNext adaptor for Illumina in one ligation reaction?
  6. Are these reagents available in a customized format, larger format or 96 well formats?
  7. Can USER Enzyme be purchased separately?
  8. Is the USER enzyme in our NEBNext kits identical in concentration to the standalone product?
  9. How do I use the NEB provided Sample Sheet for my specific instrument?
  10. Are the NEBNext Oligos for Illumina compatible with single read and paired end sequencing?
  11. What sequences need to be trimmed for NEBNext libraries that are sequenced on an Illumina instrument?

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

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

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.

Licenses


This product is covered by one or more patents.

This product is licensed from Bio-Rad Laboratories, Inc. under U.S. Pat. Nos. 6,627,424, 7,541,170, 7,670,808, 7,666,645, 7,919,296, 8,232,078, 8,367,376, 8,415,129, 8,445,249, 8,470,573, 8,476,045, 8,895,283 and 8,900,846 and corresponding patents in other countries for use only in: (a) standard (non-real time) PCR in the research field only, but not  digital PCR; (b) real-time PCR for internal product research and development purposes only, and not for sales to end-users within the research field; (c) any in vitro diagnostics application, including applications using real-time PCR but not digital PCR; and (d) any non-PCR applications in DNA sequencing, isothermal amplification and the production of synthetic DNA.

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