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.
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).
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.
The following reagents are supplied with this product:
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