Product Class:

NEBNext® Multiplex Oligos for Illumina® (Index Primers Set 1)
Item# Description List Price Web Price Qty
E7335L NEBNext Multiplex Oligos for Illumina (Index Primers 1-12) - 96 rxns $558.00
E7335S NEBNext Multiplex Oligos for Illumina (Index Primers 1-12) - 24 rxns $152.00
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at

Product Introduction

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

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.

Additional NEBNext Multiplex Oligos products are available with sets of 12 and 96 single index primers as well as dual index primers.

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

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.

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 #E7500,  NEB #E7710,  NEB #E7730,  NEB #E6609,  NEB #E7600). A methylated version of the NEBNext Adaptor is also available for use with bisulfite sequencing protocols (NEB #E7535).

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.

Figure 1: Workflow
Figure 1: Workflow.

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

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C) Concentration
NEBNext Universal PCR 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
NEBNext Adaptor for Illumina -20
USER® Enzyme -20
Product Categories:
Adaptors and Primers for Library Prep Products,
ChIP-Seq Library Prep Products,
FFPE DNA Products
Microbiome DNA Enrichment Products
DNA Library Prep Products
RNA Library Prep Products
RNA For Illumina,
Illumina Library Preparation
Applications of USER® and Thermolabile USER II Enzymes

Properties & Usage

Materials Required but not Supplied

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

    Storage Temperature


    Protocols, Manuals & Usage


    1. Refer to the manual (NEB #E7335) for a list of NEBNext library prep kits that contain protocols for use of NEBNext adaptors and primers.


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

    Faqs & Troubleshooting


    1. Are NEBNext adaptors and primers for Illumina validated in Next Generation Sequencing Workflows?
    2. Are NEBNext adaptor and primers for Illumina compatible with NEBNext reagents for Illumina library preparation?
    3. Is USER enzyme required for library preparation using NEBNext adaptor and primers for Illumina?
    4. How much DNA can I ligate to the NEBNext adaptor for Illumina in one ligation reaction?
    5. Are these reagents available in a customized format? Larger format or 96 well format?
    6. Can USER Enzyme be purchased separately?
    7. Are the NEBNext Oligos for Illumina compatible with single read and paired end sequencing?
    8. Is the USER enzyme in our NEBNext kits identical in concentration to the standalone product?
    9. How do I know the concentration of my index primers? 
    10. What should I do if the sample sheet from the website is not compatible with the software version I am using?


    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.

    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.