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NEBNext® ARTIC SARS-CoV-2 Companion Kit (Oxford Nanopore Technologies®)

Catalog # Concentration Size List Price Your Price
E7660L 96 reactions $1,188.00
$1,069.20
E7660S 24 reactions
Catalog # Size List Price Your Price
E7660L 96 reactions $1,188.00
$1,069.20
E7660S 24 reactions
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

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.

This kit includes two options for primers, including new VarSkip Short v2 primers:

  1. The new VarSkip (for Variant Skip) Short v2 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 variant overlaps can be found here.

  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. 

The Express protocol option removes clean ups of cDNA amplicons and after ligation.

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. If only the reagents for cDNA synthesis and targeted cDNA amplification are required, the NEBNext ARTIC SARS-CoV-2 RT-PCR Module (NEB #E7626) is available. Note that E7650 does not include VarSkip v2 primers.

  • Includes two primer options: VarSkip Short v2, for improved Omicron coverage, or ARTIC V3
  • Information on primer variant overlaps can be found here
  • Improved uniformity of SARS-CoV-2 genome coverage, with balanced primer pools
  • Express protocol option available, with reduced cleanup steps
  • Streamlined, high-efficiency protocol
  • Effective with a wide range of viral genome inputs (10-10,000 copies)
  • For a bulk format of the kit (2,304 reactions) please contact us

1.(Josh Quick 2020. nCoV-2019 sequencing protocol v2 (GunIt). protocols.io https://dx.doi.org/10.17504/protocols.io.bdp7i5rn).

Featured Videos
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  • Monarch_MiniprepProtocol_VideoThumb
    Improving genome coverage in SARS-CoV-2 sequencing

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.

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 reduce the impact of SARS-CoV-2 variants on amplification and provide improved performance, including with 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 found here.

  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.

The Express protocol removes the cleanup of cDNA Amplicons.

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. If only the reagents for cDNA synthesis and targeted cDNA amplification are required, the NEBNext ARTIC SARS-CoV-2 RT-PCR Module (NEB #E7626) is available.  Note that E7650 does not include VarSkip v2 primers.

Figure 1: Sequence coverage of a SARS-CoV-2 Omicron clinical sample with updated primer schemes



Caption: Integrative Genome Viewer visualization of read coverage across the SARS-CoV-2 genome (log scale). Amplicons were generated from the same Omicron variant SARS-CoV-2 viral gRNA clinical sample using NEBNext® VarSkip Short SARS-CoV-2 primer pools, NEBNext VarSkip Short v2 SARS-CoV-2 primer pools, MilliporeSigma ARTICv4, MilliporeSigma ARTICv4 primer pools plus IDT ARTICv4.1 spike-in primers (ARTICv4.1). Libraries were constructed using the NEBNext ARTIC SARS-CoV-2 Companion Kit (ONT) and sequenced on a GridION. Reads were down-sampled to 74000 reads with seqtk and aligned to SARS-CoV-2 reference genome (NCBI, NC_045512) with minimap2.



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



Caption: Integrative Genome Viewer visualization of read coverage across the SARS-CoV-2 genome (log scale). Amplicons were generated from the same Delta variant SARS-CoV-2 viral gRNA clinical sample using NEBNext® VarSkip Short SARS-CoV-2 primer pools, NEBNext VarSkip Short v2 SARS-CoV-2 primer pools, MilliporeSigma ARTICv4, MilliporeSigma ARTICv4 primer pools plus IDT ARTICv4.1 spike-in primers (ARTICv4.1). Libraries were constructed using the NEBNext ARTIC SARS-CoV-2 Companion Kit (ONT) and sequenced on GridION. Reads were down-sampled to 74000 reads with seqtk and aligned to SARS-CoV-2 reference genome (NCBI, NC_045512) with minimap2.


Figure 3: Express and Standard workflows for the NEBNext ARTIC SARS-CoV-2 Companion Library Prep Kit for Oxford Nanopore with VarSkip Short v2 or NEBNext ARTIC primers



 
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  NEBNext® ARTIC SARS-CoV-2 Companion Kit (Oxford Nanopore Technologies®) E7660-2 -20 1 x 96 reactions
  NEBNext® Sample Purification Beads E7666L 25 1 x 96 reactions
  NEBNext® ARTIC SARS-CoV-2 Primer Mixes E7871L -20 1 x 96 reactions
Features
  • 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 
  • Optional use control human primers provided
  • Includes NEBNext Sample Purification Beads (SPRIselect®)
 


Companion Products
References
  • Josh Quick (2020). nCoV-2019 sequencing protocol v2 (GunIt) . protocols.io.
FAQs
Quality Control Assay
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
Change Notifications

Effective 6/15/2022, primer components of this product are provided in a separate box.

Effective 2/15/2022 VarSkip Short v2 SARS-CoV-2 Primer Mixes 1 and 2 replace VarSkip Short SARS-CoV-2 Primer Mixes 1 and 2.

Specifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
Certificate of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Safety Data Sheets
Legal And Disclaimer

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

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.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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 busdev@neb.com.

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