Product Class: Kit

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

This product was discontinued on December 15, 2022. An alternative product is the NEBNext® Multiplex Oligos for Enzymatic Methyl-seq (Unique Dual Index Primer Pairs) (NEB #E7140).

  • Catalog # E7535 was discontinued on December 15, 2022

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 a methylated version of the NEBNext adaptor, which is compatible with library preparation for bisulfite sequencing protocols.

  • High efficiency adaptor ligation
  • Minimized adaptor-dimer formation
  • Includes index primers for library multiplexing
  • Compatible with bisulfite sequencing protocols
Bioz Badge Exists : True
Catalog # Size Concentration

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.

This product is related to the following categories:

Kit Components

Kit Components

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • E7535S     -20    
      NEBNext Methylated Adaptor for Illumina E7536AVIAL -20 1 x 0.24 ml Not Applicable
      USER® Enzyme E7338AVIAL -20 1 x 0.072 ml Not Applicable
      NEBNext Index 1 Primer for Illumina E7311AVIAL -20 1 x 0.01 ml 10 µM
      NEBNext Index 2 Primer for Illumina E7312AVIAL -20 1 x 0.01 ml 10 µM
      NEBNext Index 3 Primer for Illumina E7313AVIAL -20 1 x 0.01 ml 10 µM
      NEBNext Index 4 Primer for Illumina E7314AVIAL -20 1 x 0.01 ml 10 µM
      NEBNext Index 5 Primer for Illumina E7315AVIAL -20 1 x 0.01 ml 10 µM
      NEBNext Index 6 Primer for Illumina E7316AVIAL -20 1 x 0.01 ml 10 µM
      NEBNext Index 7 Primer for Illumina E7317AVIAL -20 1 x 0.01 ml 10 µM
      NEBNext Index 8 Primer for Illumina E7318AVIAL -20 1 x 0.01 ml 10 µM
      NEBNext Index 9 Primer for Illumina E7319AVIAL -20 1 x 0.01 ml 10 µM
      NEBNext Index 10 Primer for Illumina E7320AVIAL -20 1 x 0.01 ml 10 µM
      NEBNext Index 11 Primer for Illumina E7321AVIAL -20 1 x 0.01 ml 10 µM
      NEBNext Index 12 Primer for Illumina E7322AVIAL -20 1 x 0.01 ml 10 µM
      NEBNext Universal PCR Primer for Illumina E6861AVIAL -20 1 x 0.12 ml 10 µM
  • E7535L     -20    
      NEBNext Methylated Adaptor for Illumina E7536AAVIAL -20 1 x 0.96 ml Not Applicable
      USER® Enzyme E7338AAVIAL -20 1 x 0.288 ml Not Applicable
      NEBNext Index 1 Primer for Illumina E7311AAVIAL -20 1 x 0.04 ml 10 µM
      NEBNext Index 2 Primer for Illumina E7312AAVIAL -20 1 x 0.04 ml 10 µM
      NEBNext Index 3 Primer for Illumina E7313AAVIAL -20 1 x 0.04 ml 10 µM
      NEBNext Index 4 Primer for Illumina E7314AAVIAL -20 1 x 0.04 ml 10 µM
      NEBNext Index 5 Primer for Illumina E7315AAVIAL -20 1 x 0.04 ml 10 µM
      NEBNext Index 6 Primer for Illumina E7316AAVIAL -20 1 x 0.04 ml 10 µM
      NEBNext Index 7 Primer for Illumina E7317AAVIAL -20 1 x 0.04 ml 10 µM
      NEBNext Index 8 Primer for Illumina E7318AAVIAL -20 1 x 0.04 ml 10 µM
      NEBNext Index 9 Primer for Illumina E7319AAVIAL -20 1 x 0.04 ml 10 µM
      NEBNext Index 10 Primer for Illumina E7320AAVIAL -20 1 x 0.04 ml 10 µM
      NEBNext Index 11 Primer for Illumina E7321AAVIAL -20 1 x 0.04 ml 10 µM
      NEBNext Index 12 Primer for Illumina E7322AAVIAL -20 1 x 0.04 ml 10 µM
      NEBNext Universal PCR Primer for Illumina E6861AAVIAL -20 1 x 0.48 ml 10 µM

Properties & Usage

Materials Required but not Supplied

Compatible with the following library preparation kits:
NEBNext Ultra™ II DNA Library Prep Kit for Illumina (#E7645),
NEBNext Ultra™ DNA Library Prep Kit for Illumina (#E7370),
NEBNext DNA Library Prep Master Mix Set for Illumina (#E6040), or
NEBNext DNA Library Prep Reagent Set for Illumina (#E6000)

NEB EpiMark® Bisulfite Conversion Kit (NEB # E3318S)
NEB EpiMark Hot Start Taq DNA Polymerase (NEB # M0490S)

10 mM dNTPs
Freshly prepared 80% Ethanol
Magnetic rack or plate (e.g., NEBNext® Magnetic Separation Rack (NEB #S1515S), Alpaqua ® 96S Super Magnet Plate (#A001322), or equivalent)
DNA LoBind® Tubes (Eppendorf®)/PCR Plates
Agencourt® AMPure® XP Beads (Beckman Coulter, Inc. #A63881)
Bioanalyzer® (Agilent® Technologies, Inc.)
Nuclease Free Water
0.1X TE (or 10 mM Tris-HCl, pH 7.5-8.0)

Protocols, Manuals & Usage

Protocols

  1. Please see manual (NEB #E7535) for protocols

Manuals

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

Usage & Guidelines

FAQs & Troubleshooting

FAQs

  1. What are the major applications of NEBNext Methylated Adaptor for Illumina?
  2. Is library prep workflow different when NEBNext Methylated Adaptor is used?
  3. How is NEBNext Methylated Adaptor (NEB #E7535) different from NEBNext adaptor for Illumina (NEB #E7335 & #E7350)?
  4. How do I know the concentration of my index primers? 
  5. What should I do if the sample sheet from the NEB.com 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.

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.

Trademarks

NEW ENGLAND BIOLABS®, NEBNEXT® and EPIMARK®are registered trademarks of New England Biolabs, Inc.
NEBNEXT® Adaptors are protected under a pending patent.
USER™ and ULTRA™ are trademarks of New England Biolabs, Inc. 
USER™ is protected by U.S. patent no. 7,435,572 (New England Biolabs, Inc.)

AGENCOURT® and AMPURE® are registered trademarks of Beckman Coulter, Inc.
AGILENT® and BIOANALYZER® are registered trademarks of Agilent Technologies, Inc.
EPPENDORF® and LOBIND® are registered trademarks of Eppendorf AG.
ILLUMINA® is a registered trademark of Illumina, Inc.
QIAQUICK® and MINELUTE® are registered trademarks of Qiagen.

Other Products You May Be Interested In