NEBNext® Multiplex Oligos for Illumina® (96 Unique Dual Index Primer Pairs Set 2)
Product informationCode | Name | Size | Quantity | Price | |
---|---|---|---|---|---|
E6442S |
NEBNext Multiplex Oligos for Illumina Set 2 (96 Unique Dual Index Primer Pairs) |
96 rxns | - | Unavailable in your region | |
E6442L |
NEBNext Multiplex Oligos for Illumina Set 2 (96 Unique Dual Index Primer Pairs) |
384 rxns | - | Unavailable in your region |
NEBNext® Multiplex Oligos for Illumina® (96 Unique Dual Index Primer Pairs Set 2)
Catalog #E6442
Product Introduction
For optimal workflow flexibility, NEBNext® Multiplex Oligos are available separately from library preparation kits, providing adaptors and primers for high-yield multiplex Illumina® library production. The NEBNext Adaptor has a unique hairpin loop structure that minimizes adaptor-dimer formation, while NEBNext index PCR primers enable index (barcode) incorporation during library amplification. This second set of unique-dual indices (UDIs) includes 96 pre-mixed unique pairs of index primers (i5 and i7), packaged in a single-use 96-well plate with a pierceable foil seal.
For additional information on available indices and formats for multiplexing, please refer to the NEBNext Multiplex Oligos Selection Chart.
- Increased sample identification specificity
- Enables complete filtering of index-hopped reads
- Highly efficient adaptor ligation
- Minimized adaptor-dimer formation
- Includes index primers for library multiplexing
- This set (Set 2) may be combined with sets 1, 3, 4 and 5 for 480 barcodes
- Product Information
- Protocols, Manuals & Usage
- Tools & Resources
- FAQs & Troubleshooting
- Citations & Technical Literature
- Quality, Safety & Legal
- Other Products You May Be Interested In
Product Information
Description
NEBNext® Adaptor ligates with increased efficiency to end-repaired, dA-tailed DNA, due to its novel hairpin-loop structure. The loop includes a uracil (U), which is removed by treatment with USER® Enzyme (a mix of UDG and Endo VIII), opening the loop and making it available as a substrate for PCR. During PCR, NEBNext index primers can be incorporated, thereby enabling multiplexing. This kit includes 96 pre-mixed unique pairs of i5 and i7 index primers, packaged in a single-use 96-well plate with a pierceable foil seal.
NEBNext Oligos are for use with NEBNext products or other standard Illumina-compatible library preparation protocols. Additional options are available in unique dual barcodes (NEB #E6440), dual barcode sets (NEB #E7600, NEB #E7780), single primer sets (in 12- and 96- index formats; NEB #E7335, NEB #E7500, NEB #E7710, NEB #E7730, NEB #E6609), and an option compatible with EM-seq™ and bisulfite sequencing (NEB #E7140).
Workflow:
NEBNext Adaptors enable high-efficiency adaptor ligation and high library yields, with minimized adaptor-dimer formation, and are designed for use in library prep for DNA, ChIP DNA and RNA (but not Small RNA). A novel hairpin loop structure ligates with increased efficiency to end-repaired, dA-tailed DNA. The loop contains a U, which is removed by USER Enzyme (a combination of UDG and Endo VIII), thereby becoming a substrate for PCR, when barcodes can be incorporated by use of the NEBNext index primers. The 96 8-base index primer pairs included in this kit are pre-mixed and are packaged in a single-use 96-well plate with a pierceable foil seal.
For multiplexing with 384 indices, combine this set with NEB #E6440, NEB #E6444, and NEB #E6446.
- This product is related to the following categories:
- NEBNext® Multiplex Oligos (Adaptors & Primers) Products,
- Next Generation Sequencing Library Preparation Products,
Kit Components
Kit Components
The following reagents are supplied with this product:
NEB # | Component Name | Component # | Stored at (°C) | Amount | Concentration | |
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Properties & Usage
Protocols, Manuals & Usage
Protocols
Manuals
Usage & Guidelines
- E6442_UDI_IlluminaLRM
- Forward Strand Workflow Sample Sheet for E6442: NovaSeq 6000 with v1.0 reagent kits, MiniSeq with Rapid reagent kits, MiSeq, HiSeq 2000/2500 (paired-end flow cell), HiSeq 3000/4000 (single-read flow cell)
- Reverse Complement Workflow Sample Sheet for E6442: iSeq 100, MiniSeq with Standard reagent kits, NextSeq Systems, NovaSeq 6000 with v1.5 reagent kits, HiSeq 2000/2500 (single-read flow cell), HiSeq 3000/4000 (paired-end flow cell)
Tools & Resources
Selection Charts
FAQs & Troubleshooting
FAQs
- Are NEBNext adaptors and primers for Illumina validated in Next Generation Sequencing Workflows?
- What is the concentration of the adaptor and primers in the NEBNext Multiplex Oligos kits?
- 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)?
- How many reactions worth of primer are in each well?
- Is the NEBNext adaptor methylated?
- 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?
- Are dual indexed libraries compatible with single end sequencing?
- How many indices are available with NEBNext® Multiplex Oligos for Illumina® (96 Unique Dual Index Primer Pairs) (NEB #E6440, E6442, E6444, E6446, and E6448)?
- Can the NEBNext Multiplex Oligos for Illumina (96 Unique Dual Index Primer Pairs) (NEB #E6440, #E6442, #E6444, #E6446, and #E6448) be used to address "index hopping" with Illumina patterned flow cells?
- Where can I find the sample sheets for these Multiplex Oligos?
- Which sets of NEBNext Unique Dual Index Primer Pairs can be combined in a single Illumina sequencing run?
- How do I use the NEB provided Sample Sheet for my specific instrument?
- What sequences need to be trimmed for NEBNext libraries that are sequenced on an Illumina instrument?
Citations & Technical Literature
Citations
Additional Citations
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]- E6442L_v1_10057833
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Safety DataSheets
The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.NEBNext® Multiplex Oligos for Illumina® (96 Unique Dual Index Primer Pairs Set 2)
USER® Enzyme
NEBNext® Adaptor for Illumina®
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.
The supporting documents available for this product can be downloaded below.