Product Class: Kit

NEBNext® LV Unique Dual Index Primers Set 2B

Please note: Beginning 06/01/2026 and phasing in on a rolling basis, NEBNext Multiplex Oligos provided in a 96-well plate will be updated to Eppendorf twin.tec® PCR Plates (Eppendorf #951020401) with Agilent Pierceable Aluminum Seals (Agilent #06644-001). This product has not yet been updated. Please contact NEBNext Technical Support (info@neb.com) with any questions related to these updates.

NEBNext® LV Unique Dual Index Primers Set 2B is the new name for NEBNext Primers for Epigenetics (Unique Dual Index Set 2B); no other changes have been made to the product.

Product Introduction

NEBNext LV Unique Dual Index Primers Set 2B includes 24 unique pairs of i5 and i7 index primers optimized for low-volume (LV) NEBNext workflows, including NEBNext EM-seq™ v2, E5hmC-seq™, and NEBNext Low-bias Small RNA Library Prep Kit, with Illumina® sequencing. The primers enable incorporation of unique index pairs during library amplification and have been designed to minimize index hopping.

  • Optimized for low-volume (LV) NEBNext library prep workflows
  • Addresses the "barcode hopping" issue seen with some Illumina sequencing instruments
  • Increases sample identification specificity with unique dual index primer pairs
Bioz Badge Exists : True
Catalog # Size Concentration
E3392S 24 reactions

Product Information

Description

NEBNext LV Unique Dual Index Primers have been selected and paired for optimal multiplex index incorporation during PCR. Optimized for low-volume (LV) NEBNext workflows, including EM-seq v2 and E5hmC-seq, these primers enable multiplexing and have been designed to minimize index hopping. They are supplied as pairs of premixed i5 and i7 8-base index primers in 24 wells of a plate that has a pierceable foil seal for easy, single use.

The index sequences from the NEBNext LV Unique Dual Index Primers Set 2B are compatible with and can be combined with the other NEBNext LV Unique Dual Index Primers sets to multiplex up to 480 reactions. Note: NEBNext LV Unique Dual Index Primers Set 2A (NEB #E3390) and Set 2B are 24-index subsets of Set 2 (NEB #E3402); therefore, we do not recommend combining Set 2A and/or 2B with Set 2

Please refer to the relevant NEBNext library prep kit manual for usage instructions. Sample sheets for use with these NEBNext LV Unique Dual Index Primers can be found under "Usage Guidelines" on this product page (see below).

This product is related to the following categories:
Enzymatic Conversion for DNA Methylation Analysis,
Hydroxymethylation Detection and Analysis,
NEBNext® Multiplex Oligos (Adaptors & Primers),
Methylome Analysis,
DNA Methylation Analysis,
Next Generation Sequencing Library Preparation,

Kit Components

Kit Components

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • NEB # Stored at (°C)
    E3392S -20
    Component Name Component # Stored at (°C) Amount Concentration
    NEBNext® LV Unique Dual Index Primers Set 2B E3399AVIAL -20 1 x 5 μl/well 10 µM

Properties & Usage

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

  1. What is the difference between NEBNext® LV Unique Dual Index Primers and NEBNext LV Unique Dual Index Primers?
  2. Are Sample Sheets available for use with the NEBNext® LV Unique Dual Index Primers Sets 1-5?
  3. Are libraries that have been dual indexed using the NEBNext® LV Unique Dual Index Primers compatible with single-end sequencing?
  4. How many primer pairs are available in the NEBNext® LV Unique Dual Index Primer Pairs Sets 1-5?
  5. Can the NEBNext® LV Unique Dual Index Primers be used to address “index hopping” with Illumina® patterned flow cells?
  6. Are the NEBNext® LV Unique Dual Index Primers the same as NEBNext Multiplex Oligos for Illumina®?
  7. Are the NEBNext® LV Unique Dual Index Primers validated for the EM-seq™ v2 or E5hmC-seq™ workflows?
  8. What is the concentration of the Index Primers in NEBNext® LV Unique Dual Index Primers?
  9. How many reactions’ worth of primer are in each well of the NEBNext® LV Unique Dual Index Primers?
  10. Can I use the 96-well primer plate supplied with the NEBNext LV Unique Dual Index Primers to do my PCR reactions?
  11. Can I use the 96-well primer plate supplied with the NEBNext® LV Unique Dual Index Primers to do my PCR reactions?
  12. Do I need to spike in custom sequencing primers when sequencing libraries made with NEBNext® LV Unique Dual Index Primers on Illumina® sequencing instruments?
  13. Can I perform single read runs and still get both index sequences? Can I run both single read and paired-end recipes with dual-indexed libraries?
  14. What sequences need to be trimmed for NEBNext libraries that are sequenced on an Illumina instrument?
  15. What do I do if I received an index collision/barcode collision error?

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]

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.

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