Product Class: Kit

NEBNext® Multiplex Oligos for Enzymatic Methyl-seq (Unique Dual Index Primer Pairs)

Product Introduction

NEBNext Multiplex Oligos for Enzymatic Methyl-seq (Unique Dual Index Primer Pairs) is compatible with Illumina® sequencing and includes the NEBNext EM-seq™ Adaptor, and NEBNext PCR primers that enable incorporation of unique index pairs during library amplification.  The index primers are provided as pre-mixed unique pairs of i5 and i7 index primers, packaged in a single-use 96-well plate with a pierceable foil seal, or in individual vials.

Although optimized for the EM-seq workflow, the NEBNext EM-seq Adaptor is also compatible with bisulfite sequencing workflows.

These components are also included in the NEBNext Enzymatic Methyl-seq Kit (NEB #E7120).

  • Optimized for the EM-seq workflow
  • Compatible with bisulfite sequencing workflows
  • Addresses the “barcode hopping” issue seen with some Illumina sequencing instruments
  • Increases sample identification specificity with unique dual index primer pairs
Catalog # Size Concentration
E7140S 24.0 reactions
E7140L 96.0 reactions

Product Information

Description

NEBNext Multiplex Oligos for Enzymatic Methyl-seq (Unique Dual Index Primer Pairs) is compatible with Illumina® sequencing and includes the NEBNext EM-seq™ Adaptor, and NEBNext PCR primers that enable incorporation of unique index pairs during library amplification.

The 96-reaction size includes a single-use 96-well plate with a pierceable foil seal. Each well of the plate contains a unique pair of one i5 index primer and one i7 index primer, therefore providing 96 unique dual indices. For the 24-reaction size, indices are provided in 24 separate vials, with each vial containing a unique pair of one i5 index primer and one i7 index primer. There is no overlap between i5 and i7 index pairs provided in the 24 and 96 reaction kits. 

Although optimized for the EM-seq workflow, the NEBNext EM-seq Adaptor is also compatible with bisulfite sequencing workflows.


These components are also provided in the NEBNext Enzymatic Methyl-seq Kit (NEB #E7120).
This product is related to the following categories:
Epigenetic Analysis Products,
NEBNext® Multiplex Oligos (Adaptors & Primers) Products,
Epigenetics Products,
Next Generation Sequencing Library Preparation Products,
This product can be used in the following applications:
DNA Methylation Analysis,
Bisulfite Conversion,
Epigenetics

Kit Components

Kit Components

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • E7140S     -20    
      EM-seq™ Index Primer 1 (S) E7141AVIAL -20 1 x 0.005 ml 10 µM
      EM-seq™ Index Primer 2 (S) E7142AVIAL -20 1 x 0.005 ml 10 µM
      EM-seq™ Index Primer 3 (S) E7143AVIAL -20 1 x 0.005 ml 10 µM
      EM-seq™ Index Primer 4 (S) E7144AVIAL -20 1 x 0.005 ml 10 µM
      EM-seq™ Index Primer 5 (S) E7145AVIAL -20 1 x 0.005 ml 10 µM
      EM-seq™ Index Primer 6 (S) E7146AVIAL -20 1 x 0.005 ml 10 µM
      EM-seq™ Index Primer 7 (S) E7147AVIAL -20 1 x 0.005 ml 10 µM
      EM-seq™ Index Primer 8 (S) E7148AVIAL -20 1 x 0.005 ml 10 µM
      EM-seq™ Index Primer 9 (S) E7149AVIAL -20 1 x 0.005 ml 10 µM
      EM-seq™ Index Primer 10 (S) E7150AVIAL -20 1 x 0.005 ml 10 µM
      EM-seq™ Index Primer 11 (S) E7151AVIAL -20 1 x 0.005 ml 10 µM
      EM-seq™ Index Primer 12 (S) E7152AVIAL -20 1 x 0.005 ml 10 µM
      EM-seq™ Index Primer 13 (S) E7153AVIAL -20 1 x 0.005 ml 10 µM
      EM-seq™ Index Primer 14 (S) E7154AVIAL -20 1 x 0.005 ml 10 µM
      EM-seq™ Index Primer 15 (S) E7155AVIAL -20 1 x 0.005 ml 10 µM
      EM-seq™ Index Primer 16 (S) E7156AVIAL -20 1 x 0.005 ml 10 µM
      EM-seq™ Index Primer 17 (S) E7157AVIAL -20 1 x 0.005 ml 10 µM
      EM-seq™ Index Primer 18 (S) E7158AVIAL -20 1 x 0.005 ml 10 µM
      EM-seq™ Index Primer 19 (S) E7159AVIAL -20 1 x 0.005 ml 10 µM
      EM-seq™ Index Primer 20 (S) E7160AVIAL -20 1 x 0.005 ml 10 µM
      EM-seq™ Index Primer 21 (S) E7161AVIAL -20 1 x 0.005 ml 10 µM
      EM-seq™ Index Primer 22 (S) E7162AVIAL -20 1 x 0.005 ml 10 µM
      EM-seq™ Index Primer 23 (S) E7163AVIAL -20 1 x 0.005 ml 10 µM
      EM-seq™ Index Primer 24 (S) E7164AVIAL -20 1 x 0.005 ml 10 µM
      NEBNext® EM-seq™ Adaptor E7165AVIAL -20 1 x 0.06 ml Not Applicable
  • E7140L     -20    
      NEBNext® 96 Unique Dual Index Primer Pairs Plate (L) E7166AVIAL -20 1 x 5 μl/well 10 µM
      NEBNext® EM-seq™ Adaptor E7165AAVIAL -20 1 x 0.24 ml Not Applicable

Properties & Usage

Protocols, Manuals & Usage

Protocols

  1. Where can I find protocols for using the NEBNext® Enzymatic Methyl-seq Kit? (NEB #E7120, E7125, E7140)

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. What is provided in the NEBNext Multiplex Oligos for Enzymatic Methyl-seq?
  2. Can the NEBNext® Multiplex Oligos for Enzymatic Methyl-seq be used to address “index hopping” with Illumina patterned flow cells?
  3. How many indices are available with the 24 reaction- or 96 reaction-size of the NEBNext® Multiplex Oligos for Enzymatic Methyl-seq?
  4. Are the NEBNext® Multiplex Oligos for Enzymatic Methyl-seq the same as NEBNext® Multiplex Oligos for Illumina®?
  5. Are the EM-seq Adaptor and Index Primers validated in the EM-seq workflow?
  6. What is the concentration of the EM-seq Adaptor and Index Primers?
  7. How many reactions worth of primer are in each well/tube?
  8. Can I use the 96-well primer plate to do my PCR reactions when using the NEBNext® Multiplex Oligos for MGI® (Dual Index Primer Pairs Set 1) (NEB #E9640)?
  9. Do I need to spike in custom sequencing primers when sequencing libraries made with NEBNext® Multiplex Oligos for Enzymatic Methyl-seq on Illumina® sequencing instruments?
  10. 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?
  11. Are Sample Sheets available for use with the NEBNext® Multiplex Oligos for EM-seq?
  12. Can the EM-seq Adaptor be used for bisulfite sequencing?
  13. Are dual indexed libraries compatible with single end sequencing?
  14. 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.

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