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.
For additional information on available indices and formats for multiplexing, please refer to the NEBNext Multiplex Oligos Selection Chart.
Note: NEBNext® Multiplex Oligos for Illumina® (Index Primer Set 1) (NEB #7335S) includes 12 indices.
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.
Additional single- or dual-barcode primer options are available, in 12- and 96- index formats (NEB #E7335, NEB #E7710, NEB #E7730, NEB #E6609, NEB #E7600). A methylated version of the NEBNext Adaptor is also available for use with bisulfite sequencing protocols (NEB #E7535).
Each set of reagents is functionally validated together through construction and sequencing of genomic DNA libraries on the Illumina sequencing platform.
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