NEBNext ARTIC SARS-CoV-2 kits are also available for smaller (~150 bp) (NEB #E7658) and larger (~400 bp) (NEB #E7650), insert libraries for Illumina sequencing.
Catalog #E7660
The NEBNext® ARTIC kits were developed in response to the critical need for reliable and accurate methods for SARS-CoV-2 sequencing. These kits, for long and short read sequencing, were based on the original work of the ARTIC Network(1). The ARTIC SARS-CoV-2 sequencing workflow is a multiplexed amplicon-based whole-viral-genome sequencing approach.
NEBNext ARTIC kits include primers and reagents for RT-PCR from SARS-CoV-2 gRNA and downstream library preparation for Illumina® and Oxford Nanopore Technologies® sequencing. The balanced V3 primers and optimized reagents for RT-PCR deliver improved uniformity of amplicon yields from gRNA across a wide copy number range.
The NEBNext ARTIC SARS-CoV-2 Companion Kit (Oxford Nanopore Technologies®) is compatible with sequencing on the Oxford Nanopore Technologies (ONT) platform and is used alongside kits from ONT to generate libraries with inserts in the 400 bp range, that can be sequenced on any ONT instrument, in 2-24 hour run times.
If Illumina-compatible libraries are preferred, the NEBNext ARTIC SARS-CoV-2 FS Library Prep Kit (Illumina) (NEB #E7658) and the NEBNext ARTIC SARS-CoV-2 Library Prep Kit (Illumina) (NEB #E7650) generate libraries with ~150 bp and ~400 bp inserts, respectively.
Streamlined, high-efficiency protocol
Effective with a wide range of viral genome inputs (10-10,000 copies)
Improved uniformity of SARS-CoV-2 genome coverage depth with a more balanced primer pool
Library inserts in the 400 bp range
Includes NEBNext Sample Purification Beads (SPRIselect®)
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 Library Prep Kit is based on this method and incorporates optimized and novel reagents.
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. The multiplex primers and PCR reaction conditions are balanced to provide uniform amplicon coverage across the viral genome.
The NEBNext ARTIC SARS-CoV-2 Companion Kit (Oxford Nanopore Technologies) is compatible with sequencing on the Oxford Nanopore Technologies platform and is used alongside kits from Oxford Nanopore Technologies to generate libraries with inserts in the 400bp range, that can be sequenced on any Oxford Nanopore Technologies instrument, in 2-24 hour run times.
If Illumina-compatible libraries are preferred, the NEBNext ARTIC SARS-CoV-2 FS Library Prep Kit (Illumina) (NEB #E7658) and the NEBNext ARTIC SARS-CoV-2 Library Prep Kit (Illumina) (NEB #E7650) generate libraries with ~150 bp and ~400 bp inserts, respectively.
Integrative Genome Viewer visualization of read coverage across the SARS-CoV-2 genome. Amplicons were generated from 1,000 copies of SARS-CoV-2 viral gRNA inputs (ATCC VR-1986 and VR-1991) in 100 ng of Universal Human Reference RNA (ThermoFisher QS0639) using IDT ARTIC nCoV-2019 V3 Panel (“Standard”) or the NEBNext balanced ARTIC SARS-CoV-2 primer pools. Libraries were constructed using the NEBNext ARTIC SARS-CoV-2 Companion Kit (Oxford Nanopore Technologies) and the Oxford Nanopore Technologies Native Barcoding Expansion kits 1-12 (EXP-NBD104) and 13-24 (EXP-NBD114), Ligation Sequencing Kit (SQK-LSK109) and SFB Expansion Kit (EXP-SFB001). Sequencing was on a GridION instrument using R9.4.1 flow cells, and the ARTIC Network nCoV-19 bioinformatics pipeline was used.
Figure 2: Improved genome coverage with the NEBNext ARTIC SARS-CoV-2 Companion Kit (Oxford Nanopore Technologies)
Amplicons were generated from 1,000 copies of SARS-CoV-2 viral gRNA inputs (ATCC VR-1991) in 100 ng of Universal Human Reference RNA (ThermoFisher QS0639) using the IDT ARTIC nCoV-2019 V3 Panel or the NEBNext balanced ARTIC SARS-CoV-2 primer pools and the standard or NEBNext protocol, as indicated. Libraries were constructed using the NEBNext ARTIC SARS-CoV-2 Companion Kit (Oxford Nanopore Technologies) and the Oxford Nanopore Technologies Native Barcoding Expansion kits 1-12 (EXP-NBD104) and 13-24 (EXP-NBD114), Ligation Sequencing Kit (SQK-LSK109) and SFB Expansion Kit (EXP-SFB001). Sequencing was on a GridION using R9.4.1 flow cells. Reads were down-sampled (seqtk) to the indicated numbers and ARTIC Network vCOV-19 bioinformatics pipeline was used to generate consensus genomes. Quast was used to compare the genome consensus to the reference SARS-CoV-2 sequence to obtain genome coverage percentage. With 100x coverage, only the most 5´ and 3´ region (~120 nucleotides) are missing from the consensus.
Figure 3: Robust SARS-CoV-2 RT-PCR amplification
Amplicons were generated from 10-10,000 copies of SARS-CoV-2 viral gRNA inputs (ATCC VR-1986 and VR-1991) in 100 ng of Universal Human Reference RNA (ThermoFisher QS0639) using NEBNext balanced ARTIC SARS-CoV-2 primer pools. Mean amplicon yield was measured using the Qubit™ dsDNA BR Assay Kit (ThermoFisher Q32850).
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This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
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Licenses
This product is covered by one or more patents.
This product is licensed for research and commercial use from Bio-Rad Laboratories, Inc., under U.S. Pat. Nos. 6,627,424, 7,541,170, 7,670,808, 7,666,645, and corresponding patents in other countries. No rights are granted for use of the product for Digital PCR or real-time PCR applications, with the exception of quantification in Next Generation Sequencing workflows. For additional information or to inquire about commercial use, please contact [email protected].
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