The NEBNext ARTIC kits meet the increasing need for reliable, accurate and fast methods for sequencing SARSCoV-2, with Oxford Nanopore Technologies or Illumina sequencing. The kits use the ARTIC multiplexed ampliconbased whole-viral-genome sequencing approach and were developed in collaboration with the ARTIC Network (artic. network). The kits include balanced V3 primer pools for improved uniformity of genome coverage and are effective with 10-10,000 SARS-CoV-2 genome copies. The reagents and protocols for RT-PCR and library prep are all optimized specifically for the SARS-CoV-2 ARTIC workflows.
New Primer Set included for better coverage of SARS-CoV-2 variants as well as new Express protocols
Starting September 2021, two options for primers are now available, 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):
- 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.
- 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 and Delta variants. The Omicron variant can be called confidently using NEBNext VarSkip Short (VSS) primers. Note that there are two dropouts (amplicons 56 and 67), and two amplicons (20 and 64) have lower coverage. Making 2 libraries, with both ARTIC V3 and VSS, will provide near complete genome coverage. The primer pools are balanced for coverage uniformity.
Information on primer overlaps with Omicron and Delta variants.
The two kits compatible with Illumina sequencing generate library inserts of ~150 bp (for 2 x 75 sequencing) or ~400 bp (for 2 x 250 sequencing), and include a novel DNA polymerase formulation that eliminates the need to normalize amplicon concentrations prior to library preparation.
- Improved uniformity of SARS-CoV-2 genome coverage
- Effective with a wide range of viral genome inputs (10–10,000 copies)
- Single RT conditions for all input amounts
- No amplicon normalization prior to library preparation (Illumina-compatible kits)
- NEBNext Sample Purification Beads (SPRIselect®) provided
NEBNext ARTIC Kit Options:
NEBNext ARTIC Kit Workflow:
NEBNext ARTIC Kit Genome Coverage:
Integrative Genome Viewer visualization of read coverage across the SARS-CoV-2 genome show the superior genome coverage and uniformity using the NEBNext primers, reagents and protocols. For detailled experimental data, please visit www.neb.com/E7660
„We have been impressed by the great performance of the NEBNext ARTIC FS kit in our sequencing of human samples containing SARS-CoV-2 RNA. With this kit we have been successfully processing samples of varying quality and quantity and have been amazed by its sensitivity and remarkable uniformity of the viral genome coverage. The quick turnaround time of the robust workflow has been truly beneficial for us.“
Dr. Vladimir Benes, Head of GeneCore EMBL Heidelberg, Early-Access-User
Improved sequence coverage of SARS-CoV-2 Delta clinical samples with VarSkip Short primers:
Consensus sequences classified as B.1.617.2 (Delta) in Massachusetts (where VarSkip Short was field tested) were collected from NBCBI Virus  and partitioned into 2,613 ARTIC V3 and 1,319 VarSkip Short sequences using the SRA “design” field (NEB_VarSkip_v1 = VarSkip Short, others assumed to be ARTIC V3). Reads were aligned to the NC_045512.2 reference using minimap2 (minimap2 -x asm5) and visualized using IGV. Consensus sequences generated from VarSkip amplicons suffered fewer dropouts. For detailled experimental data, please visit www.neb.com/E7658
As of: 14.05.2021