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

Especially with the ongoing emergence of SARS-CoV-2 variants that affect virus transmission and other metrics significant to public health, there is an increasing need for reliable, accurate and fast methods for sequencing SARS-CoV-2. The NEBNext ARTIC kits are based on the original work of the ARTIC Network, who quickly adapted their protocols to SARS-CoV-2 (Josh Quick 2020. nCoV-2019 sequencing protocol v2 (GunIt)).

The ARTIC method is a multiplexed amplicon-based whole-viral-genome sequencing approach (Figure 1), and the NEBNext ARTIC kit options are compatible with Illumina and Oxford Nanopore Technologies sequencing platforms. The two kits compatible with Illumina sequencing generate library inserts of ~150 bp or ~400 bp, for 2 x 75 or 2 x 250 sequencing, respectively (Figure 2). The NEBNext ARTIC SARS-CoV-2 RT-PCR Module contains only the reagents required for cDNA synthesis and targeted cDNA amplification from SARS-CoV-2 genomic RNA. 

Express workflow options are provided, with reduced cleanup steps.

Two options for primers are now available, including new VarSkip Short v2 primers, in the NEBNext ARTIC SARS-CoV-2 FS Library Prep Kit (Illumina) (NEB #E7658), NEBNext ARTIC SARS-CoV-2 Companion Kit (Oxford Nanopore Technologies) (NEB #E7660) and NEBNext ARTIC SARS-CoV-2 RT-PCR Module (NEB #E7626).

1. The VarSkip (for Variant Skip) Short primers have been designed at NEB to reduce the impact of SARS-CoV-2 variants on amplification and provide improved performance, including with the Omicron variant. The primer pools are balanced for coverage uniformity. 
   
   Information on primer overlaps with Omicron and Delta variants.
   
   
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. 

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Figure 1: NEBNext ARTIC Standard Workflow Overview. Express protocols are also available, with fewer clean ups.

Figure 2: NEBNext ARTIC Kit Options

Figure 3: Sequence coverage of SARS-CoV-2 genomes with VarSkip Short v2 plus spike-in primers 

Figure 4: Coverage of a SARS-CoV-2 Omicron clinical sample with updated primer schemes and Oxford Nanopore Technologies sequencing

RT reaction conditions are the same for all input amounts, and for Illumina applications, a novel DNA polymerase formulation for the enrichment of next-generation sequencing libraries eliminates the need to normalize amplicon concentrations prior to library preparation.

• Two primer options included: VarSkip Short v2, for improved Omicron coverage, and ARTIC V3
• Express protocols available, with reduced cleanup steps
• Improved uniformity of SARS-CoV-2 genome coverage depth with balanced primer pools
• Streamlined, high-efficiency protocols
• Effective with a wide range of viral genome inputs (10-10,000 copies)
• Available for Illumina and Oxford Nanopore Technologies sequencing platforms
• Single RT conditions for all input amounts
• No requirement for amplicon normalization prior to library preparation (Illumina-compatible kits)
• Optional use control human primers provided
• Includes NEBNext Sample Purification Beads (SPRIselect^®)
• Library adaptors and primers available separately
• Bulk formats (2,304 reactions) also available. Please contact us