Product Class: Kit

NEBNext® Multiplex Oligos for Illumina® (Dual Index Primers Set 1)

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. This kit includes 8 i5 index primers and 12 i7 index primers for dual indexing.

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
  • Increased sample identification specificity
Catalog # Size Concentration
E7600S 96.0 reactions
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Product Information


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. This kit includes 8 i5 index primers and 12 i7 index primers.

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

Single primer options are also available, in 12- and 96- index formats (NEB #E7335, NEB #E7500, NEB #E7710, NEB #E7730, NEB #E6609). A methylated version of the NEBNext Adaptor is also available for use with bisulfite sequencing protocols (NEB #E7535).

Dual Index Workflow
Dual Index Workflow

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
This product is related to the following categories:
NEBNext® Multiplex Oligos (Adaptors & Primers)
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
  • E7600S     -20    

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


  1. Setting up the PCR Reaction - NEBNext Multiplex Oligos for Illumina (Dual Index Primers Set 1) (E7600)


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


  1. Where can I find the sample sheets for these Multiplex Oligos?
  2. Do NEBNext Multiplex Oligos for Illumina (Dual Index Primers Set 1)come with library prep reagents?
  3. Can I use NEBNext Multiplex Oligos for Illumina (Dual Index Primers Set 1) for library prep from both DNA and RNA samples?
  4. How many libraries can I prepare with the NEBNext Multiplex Oligos for Illumina (Dual Index Primers Set 1)?
  5. For pooling <96 samples, what combination of indices should I use?
  6. Can I perform single read runs and still get both index sequences? 
  7. Are dual indexed libraries compatible with single end sequencing?
  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. 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.


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

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.