NEBNext® Varskip Short ARTIC SARS-CoV-2 FS Library Prep Kit (Illumina®) E7658 Express Protocol with One Clean-up Step v1
This protocol details methods for the NEBNext® ARTIC SARS-CoV-2 FS Library Prep Kit (Illumina®), NEB #E7658S/L 24/96 reactions. This protocol follows an alternate variant-tolerant approach for targeting SARS-CoV-2 by utilizing NEBNext VarSkip Short SARS-CoV-2 Primer Mixes. The NEBNext VarSkip Short SARS-CoV-2 Primer mixes cannot be added to the same cDNA amplification reaction as the NEBNext ARTIC SARS-CoV-2 Primer Mixes. This protocol does not include a cleanup step for each sample after cDNA synthesis and after adaptor ligation. Performing RNA input normalization prior to cDNA synthesis and targeted amplification and/or normalizing final libraries prior to sequencing promotes more even distribution of reads across libraries. Skipping RNA input normalization, final library normalization, and cleanups reduces hands on time but may require deeper sequencing depth to reach sufficient coverage of each sample. For other NEBNext® ARTIC SARS-CoV-2 protocols, please see the NEBNext ARTIC Protocols Collection.