Adaptors and Primers are an essential component of your NGS sample prep workflow, and NEBNext Multiplex Oligos offer flexibility in multiplexing; indexing options include unique dual indices (UDIs) with unique molecular identifiers (UMIs), unique dual indices (UDIs), combinatorial dual (CD) indices, and single indices in a range of formats and indexing strategies. For an overview of our Multiplex Oligos products, refer to the NEBNext Multiplex Oligos Selection Chart.
NEBNext index primers are intended to be combined with one of the NEBNext adaptor types. The NEBNext Adaptor, with its unique hairpin-loop shape, was designed to minimize the formation of adaptor-dimers during the adaptor-ligation step, whereas the truncated EM-seq™ Adaptor is optimized for use with Enzymatic Methyl-seq workflows. The NEBNext SR Adaptor is used in preparation of NEBNext Small RNA libraries.
NEBNext index adaptors are available for either DNA-seq or RNA-seq workflows, and these full-length index adaptors include UMIs to enable increased sample identification specificity and more-sensitive detection of low-frequency single nucleotide variants (SNVs). UMIs also allow for PCR-free DNA-seq workflows, to mitigate the risk of bias introduced by PCR cycles.
NEBNext Multiplex Oligos currently enable multiplexing of up to 96 unique dual indexed UMI reactions, up to 384 unique dual indexed reactions, up to 384 dual indexed reactions (using combinatorial indexing), and up to 144 single indexed reactions.
NEBNext Multiplex Oligos are supplied separately from NEB Next Library Prep Kits (except for the Small RNA and EM-seq kits) to provide maximal workflow flexibility.
- Are the NEBNext Oligos for Illumina compatible with single read and paired end sequencing?
- Are NEBNext adaptor and primers for Illumina compatible with NEBNext reagents for Illumina library preparation?
- What is the concentration of the adaptor and primers in the NEBNext Multiplex Oligos kits?
- Are NEBNext adaptor and primers for Illumina validated in Next Generation sequencing workflows?
- Can the NEBNext® Multiplex Oligos for Illumina® (96 Unique Dual Index Primer Pairs) (NEB #E6440, E6442, E6444, and E6446) be used to address “index hopping” with Illumina patterned flow cells?
- Is the NEBNext adaptor methylated?
With NEBNext Multiplex Oligos, you can expect the following benefits:
- Indexing strategies optimized by application
- Index Primers – For NGS Library Prep workflows that include an amplification step
- Index Adaptors – Enablement of PCR-free workflows and incorporation of UMIs for error correction/deduplication
- Extensively QC’d for purity and increased library yields
- Flexibility for use with NEBNext library preparation kits and other standard, Illumina-compatible library preparation methods
- Provided with index-pooling guidelines and sample sheets
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