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NEBNext® Multiplex Oligos for Illumina® (96 Index Primers)
Item# Description List Price Web Price Qty
E6609L NEBNext Multiplex Oligos for Illumina (96 Index Primers) 384 rxns $3,692.00
$3,322.80
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E6609S NEBNext Multiplex Oligos for Illumina (96 Index Primers) 96 rxns $923.00
$830.70
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*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

    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.
    The 96 8-base index primers included in this kit are pre-mixed with the universal primer and are packaged in a single-use 96-well plate with a pierceable foil seal.

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

    • High efficiency adaptor ligation
    • Minimized adaptor-dimer formation
    • Includes index primers for library multiplexing
    • 96-well plate format with pierceable foil seal

    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.

    The 96 8-base index primers included in this kit are pre-mixed with the universal primer and are packaged in a single-use 96-well plate with a pierceable foil seal.

    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 #E7335, NEB #E7500, NEB #E7710, NEB #E7730, 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 demonstrating the use of NEBNext Multiplex Oligos for Illumina (96 Index Primers)



    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
    USER® Enzyme -20
    NEBNext® Index/Universal Primer Mix (1-96) -20
    NEBNext® Adaptor for Illumina® -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
    Applications:
    DNA Library Preparation,
    Sample Prep for NGS & Target Enrichment,
    Illumina Library Preparation
    ,
    Applications of USER® and Thermolabile USER II Enzymes

    Properties & Usage

    Materials Required but not Supplied

    • Enzymes and buffers appropriate for DNA, ChIP or mRNA Library Preparation (NEB #E7645, E7595, E7420, E7530, E7370, E7445, E6040, E6110, E6240, E6056, E6000, E6200, E6100)
    • SPRIselect or AMPure® XP Beads (Beckman Coulter, Inc.)
    • DNA LoBind® Tubes (Eppendorf®)/PCR Plates
    • Magnetic Stand
    • Bioanalyzer® (Agilent Technologies, Inc.)
    • Freshly Prepared 80% Ethanol

    Please Refer to the Kit Specific Protocol for using the NEBNext Multiplex Oligos for Illumina:
    The following kits are designed for use with the NEBNext Multiplex Oligos for Illumina:

    • #E7760, NEBNext Ultra II Directional RNA Library Prep Kit for Illumina
    • #E7765, NEBNext Ultra II Directional RNA Library Prep with Sample Purification Beads
    • #E7770, NEBNext Ultra II RNA library Prep Kit for Illumina
    • #E7775, NEBNext Ultra II RNA Library Prep with Sample Purification Beads
    • #E7805, NEBNext Ultra II FS DNA Library Prep Kit for Illumina
    • #E6177, NEBNext Ultra II FS DNA Library Prep Kit with Sample Purification Beads
    • #E7645, NEBNext Ultra II DNA Library Prep Kit for Illumina
    • #E7103, NEBNext Ultra II DNA Library Prep with Sample Purification Beads
    • #E7595, NEBNext Ultra II Ligation Module
    • #E7420, NEBNext Ultra Directional RNA Library Prep Kit for Illumina
    • #E7530, NEBNext Ultra RNA Library Prep Kit for Illumina
    • #E7370, NEBNext Ultra DNA Library Prep Kit for Illumina
    • #E7445, NEBNext Ultra Ligation Module
    • #E6040, NEBNext DNA Library Prep Master Mix Set for Illumina
    • #E6240, NEBNext ChIP-Seq Library Prep Master Mix Set for Illumina
    • #E6056, NEBNext Quick Ligation Module

    Protocols, Manuals & Usage

    Protocols

    1. Setting up the PCR Reactions
    2. Index Pooling Guidelines - 96 Single Index Kit (E6609)

    Manuals

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

    Usage & Guidelines

    • Sample Sheet with the index sequences that can be used in the Illumina Experiment Manager

    Faqs & Troubleshooting

    FAQs

    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. Are the NEBNext Oligos for Illumina compatible with single read and paired end sequencing?
    4. What is the concentration of the adaptor and primers in the NEBNext Multiplex Oligos kits?
    5. Can I use the 96-well primer plate to do my PCR reactions?
    6. How many reactions worth of primer are in each well?
    7. Do you have a Sample Sheet with the index sequences that can be used in the Illumina Experiment Manager (IEM)? 
    8. Is the adaptor methylated?
    9. Is the USER enzyme in our NEBNext kits identical in concentration to the standalone product?
    10. Is USER enzyme required for library preparation using NEBNext adaptor and primers for Illumina?
    11. Can USER Enzyme be purchased separately?
    12. Are these reagents available in a customized format?
    13. What should I do if the sample sheet from the NEB.com website is not compatible with the software version I am using?

    Troubleshooting

    NEBNext® DNA Library Prep Reagent Set for Illumina®

    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

    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.

    Trademarks

    NEW ENGLAND BIOLABS® and NEBNEXT® are registered trademarks of New England Biolabs, Inc.
    USER is a trademark of New England Biolabs, Inc.
    AMPURE® is a registered trademark of Beckman Coulter, Inc.
    BIOANALYZER® is a registered trademark of Agilent Technologies, Inc.
    EPPENDORF® and LOBIND® are registered trademarks of Eppendorf AG.
    ILLUMINA®, HISEQ® and MISEQ® are registered trademarks of Illumina, Inc.

    © Copyright 2016, New England Biolabs, Inc; all rights reserved.