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 #E7500, NEB #E7710, NEB #E6609, NEB #E7600). A methylated version of the NEBNext Adaptor is also available for use with bisulfite sequencing protocols (NEB #E7535).
Figure 1: Workflow demonstrating the use of NEBNext Multiplex Oligos for Illumina (Index Primers Set 4)
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