New England Biolabs (NEB®) today released three kits based on the ARTIC Network protocols for multiplexed amplicon-based sequencing of viral RNA: NEBNext® ARTIC SARS-CoV-2 Library Prep Kit and NEBNext ARTIC SARS-CoV-2 FS Library Prep Kit, both for Illumina sequencing; and NEBNext ARTIC SARS-CoV-2 Companion Kit for Oxford Nanopore Technologies sequencing. The kits offer streamlined, high-efficiency protocols with balanced V3 primers and optimized reagents for uniform viral genome coverage across a range of inputs.
The ARTIC Network, an international consortium of researchers, has developed protocols for the epidemiological monitoring of viral genomes to provide insights into virus evolution and transmission for multiple viral outbreaks. Dr. Joshua Quick of the University of Birmingham in the United Kingdom, has led the development of protocols applicable to the SARS-CoV-2 outbreak based on amplicon sequencing-based approaches previously used for Ebola and Zika virus.
“Since the onset of the COVID-19 pandemic, the ARTIC protocols have been widely adopted by researchers for sequencing the SARS-CoV-2 genome, and more recently have played a key role in helping to track the emergence of new variants,” said Fiona Stewart, Ph.D., Product Portfolio Manager, Next Generation Sequencing at NEB. “Working closely with the ARTIC Network, we developed the NEBNext ARTIC SARS-CoV-2 kits for labs with all levels of experience sequencing SARS-CoV-2. The uniformity of genome coverage from the balanced primers enables rapid, cost-effective SARS-CoV-2 sequencing.”
All three of the NEBNext ARTIC SARS-CoV-2 kits deliver uniform, ample amplicon yields from gRNA across a wide copy number range and use the same RT step for all inputs. The two library prep kits for Illumina sequencing provide options for library insert size and downstream sequencing parameters.
Vladimir Benes, Head of Genomics Core Facility at the European Molecular Biology Laboratory commented “We have been impressed by the great performance of the NEBNext ARTIC SARS-CoV-2 FS Library Prep Kit (Illumina) 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.”
“The NEBNext ARTIC SARS-CoV-2 Companion Kit (Oxford Nanopore Technologies) will provide researchers with the most convenient way to perform nanopore sequencing of SARS-CoV-2,” said Joshua Quick, Ph.D., of the University of Birmingham and the ARTIC Network.
“When developing the ARTIC protocol we have focused on recovery of near-complete genomes from challenging clinical samples with < Ct33. We adopted a 400 bp tiling amplicon approach because it provides robust performance across a wide range of inputs including high Ct and degraded RNA samples which are often encountered. The native barcoding workflow provides full-length amplicons compatible with best practice demultiplexing and reference-based consensus building while the PCR-free library preparation minimizes amplicon contamination risk. We have worked in collaboration with NEB in optimizing the protocol to minimize reagent volumes while improving performance and this now also benefits from optimized primer pools which greatly improve genome completeness and reduce read coverage requirements. The compatibility with the 96-barcode native barcoding expansion kit allows the flexibility to run between 24 and 96 samples depending on your requirements.”
Source: Company Press Release