NEBNext® Multiplex Oligos for Illumina® (96 Index Primers)

Description

Designed for use in library prep for DNA, ChIP DNA and RNA (but not Small RNA), the NEBNext Adaptors enable high-efficiency adaptor ligation and high library yields, with minimized adaptor-dimer formation. 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 combination 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.


Figure 1:
Workflow demonstrating the use of NEBNext Multiplex Oligos for Illumina (96 Index Primers)



Functionally Validated: Each set of reagents is functionally validated together through construction and sequencing of genomic DNA libraries on the Illumina platform.

Lot Control: The lots provided in the NEBNext Multiplex Oligos for Illumina (96 Index Primers) are managed separately and are qualified by additional functional validation. Individual reagents undergo standard enzyme activity and quality control assays, and also meet stringent criteria in the additional quality controls listed on each individual component page.

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C)Concentration
NEBNext® Adaptor for Illumina®-20
USER Enzyme-20
NEBNext® Index/Universal Primer Mix (1-96)-20

Properties and 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:

  • #E7645 , NEBNext Ultra II DNA Library Prep Kit for Illumina
  • #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
  • #E6110 , NEBNext mRNA Library Prep Master Mix Set for Illumina
  • #E6240 , NEBNext ChIP-Seq Library Prep Master Mix Set for Illumina
  • #E6056 , NEBNext Quick Ligation Module
  • #E6000 , NEBNext DNA Library Prep Reagent Set for Illumina
  • #E6200 , NEBNext ChIP-Seq Library Prep Reagent Set for Illumina
  • #E6100 , NEBNext mRNA Library Prep Reagent Set for Illumina

Troubleshooting

Troubleshooting

NEBNext® DNA Library Prep Reagent Set for Illumina®

FAQs

  1. Are NEBNext adaptor 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 end 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?

Protocols

  1. Setting up the PCR Reactions < 96 Samples and 96 Samples - 96 Single Index Kit (E6609)
  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. The following file naming structure is used to name these document files: manual[Catalog Number].

Usage Guidelines & Tips

Safety Data Sheet

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.