The ARTIC SARS-CoV-2 protocol from the ARTIC Network is a multiplexed amplicon approach covering the whole viral genome. The NEBNext ARTIC SARS-CoV-2 RT-PCR Module is based on this method and incorporates optimized reagents for the cDNA synthesis and targeted cDNA amplification steps.A single RT protocol is used regardless of input amount (10-10,000 viral genome copies), and no normalization step is required ahead of targeted amplification.This kit includes two options for primers, including new VarSkip Short v2 primers:

1. The VarSkip (for Variant Skip) Short primers have been designed at NEB to provide improved performance with SARS-CoV-2 variants, including the Omicron variant. VarSkip Short v2 primer pools generate ~550bp amplicons. Please see the FAQ section and the product manual for additional detail.Information on primer variant overlaps can be foundhere.
2. The V3 ARTIC primers have been balanced, using methodology developed at NEB based on empirical data from sequencing. In combination with optimized reagents for RT-PCR, the kits deliver improved uniformity of amplicon yields from gRNA across a wide copy number range.

Full kits that also include library prep reagents optimized for the ARTIC workflow are available, for smaller (~150 bp) (NEB #E7658) or larger (~400 bp) (NEB #E7650) insert libraries for Illumina® sequencing, and for Oxford Nanopore Technologies^® sequencing (NEB #E7660).Note that E7650 does not include VarSkip v2 primers.

Figure 1: Sequence coverage of SARS-CoV-2 genomes with VarSkip Short v2 plus spike-in primers

Figure 2: Sequence coverage of a SARS-CoV-2 Omicron clinical sample with updated primer schemes

Figure 3: Workflow for the NEBNext ARTIC SARS-CoV-2 RT-PCR Module with VarSkip Short v2 or NEBNext ARTIC primers

This product is related to the following categories:

NEBNext® ARTIC products for SARS-CoV-2 sequencing,

RNA Library Prep for Illumina

This product can be used in the following applications:

RNA-seq,

RNA Analysis