NEBNext® Multiplex Oligos for Illumina® (96 Unique Dual Index Primer Pairs)
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. This first set of UDI primer pairs (Set 1 of 5) 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® 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.
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Additional Citations
Butler, D.J. et al. (2020) Shotgun transcriptome and isothermal profiling of SARS-CoV-2 infection reveals unique host responses, viral diversification, and drug interactions bioRxiv; DOI: https://doi.org/10.1101/2020.04.20.048066
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