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NEBNext® ARTIC SARS-CoV-2 FS Library Prep Kit (Illumina®)

Catalog # Concentration Size List Price Quantity Your Price
E7658L 96 reactions $2,414.00
$2,172.60
E7658S 24 reactions $603.00
$542.70
Catalog # Size List Price Your Price
E7658L 96 reactions $2,414.00
$2,172.60
E7658S 24 reactions $603.00
$542.70
Catalog #
Qty:
 
*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, for improved Omicron coverage:

  1. See below, including Figure 1 in the Product Information section, for information on VarSkip Short v2 performance with spike-in primers. 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. 

An Express protocol option with fewer cleanups is available.

The NEBNext ARTIC SARS-CoV-2 FS Library Prep Kit (Illumina) incorporates enzymatic cDNA fragmentation and produces Illumina-compatible libraries with library inserts in the 150 bp range, compatible with 2 x 75 sequencing on Illumina instruments.

  • Includes two primer options: new VarSkip Short v2, for improved Omicron coverage, and 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)
  • No requirement for amplicon normalization prior to library preparation
  • Library inserts in the 150 bp range
  • Compatible with 2 x 75 Illumina sequencing
  • Optional use control human primers provided
  • Includes NEBNext Sample Purification Beads (SPRIselect®)
  • Adaptors and primers available separately; For a 384 unique dual index format, please contact us
  • For a bulk format of the kit (2,304 reactions) please contact us

If larger inserts are preferred, the NEBNext ARTIC SARS-CoV-2 Library Prep Kit (Illumina) (NEB #E7650) generates libraries with ~400 bp inserts, compatible with 2 x 250 bp sequencing.

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

 
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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 FS 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, for improved Omicron coverage

  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. 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.

Library preparation uses the NEBNext FS reagents and workflow, incorporating enzymatic cDNA fragmentation, and reagent volumes tailored to the ARTIC application. Note that adaptors and primers for library prep are purchased separately. The novel NEBNext Library PCR Master Mix allows use of the same number of PCR cycles to amplify all libraries, regardless of initial input amounts. 

The Express protocol option removes the cleanup of cDNA amplicons and the cleanup of adaptor-ligated DNA. The Express protocol is recommended for inputs of ≥ 100 copies of the (SARS-CoV-2) viral genome.

The NEBNext ARTIC SARS-CoV-2 FS Library Prep Kit generates libraries with inserts ~150 bp. If longer inserts are preferred, the NEBNext ARTIC SARS-CoV-2 Library Prep Kit (NEB E7650) produces ~400 bp insert libraries, compatible with 2 x 250 Illumina sequencing. Note that this kit does not contain VarSkip v2 primers. For Oxford Nanopore Technologies® sequencing the NEBNext ARTIC SARS-CoV-2 Companion Kit (Oxford Nanopore Technologies) (NEB #E7660) can be used. 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.

 

Figure 1: Sequence coverage of SARS-CoV-2 genomes with VarSkip Short v2 plus spike-in primers 



Consensus genomes known to be generated using VarSkip Short 2 (+ supplementary primers) (green) are compared with other genomes (yellow) submitted to Genbank between 2022-08-01 and 2022-09-24. Genomes were aligned to the NC_045512.2 reference (minimap2 -x map-ont -r 20000 --score-N=0, v.2.24). Ns were tabulated using Qualimap (v2.2.2).



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



Integrative Genome Viewer visualization of read coverage across the SARS-CoV-2 genome (log scale). Amplicons were generated using NEBNext® VarSkip Short SARS-CoV-2 v2 primer pools, NEBNext VarSkip Short SARS-CoV-2 primer pools, NEBNext ARTIC SARS-CoV-2 primer pools (ARTICv3), MilliporeSigma ARTICv4 primer pools plus IDT ARTICv4.1 spike-in primers (ARTICv4.1), or MilliporeSigma ARTICv4 primer pools. The same Omicron variant SARS-CoV-2 viral gRNA clinical sample served as a template for all primer schemes except ARTICv3. Libraries were constructed using the NEBNext ARTIC SARS-CoV-2 FS Library Prep Kit (Illumina®) and sequenced on a MiSeq® instrument (2x75 bp). Coverage depth per base was determined, reads were down-sampled to 0.5M PE reads with seqtk and aligned to SARS-CoV-2 reference genome (NCBI, NC_045512) with Bowtie2.




Figure 3: Express and Standard workflows for the NEBNext ARTIC SARS-CoV-2 FS Library Prep Kit for Illumina 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 FS Library Prep Kit (Illumina®) E7658-1 -20 1 x 24 reactions
  NEBNext® Sample Purification Beads E7659S 25 1 x 24 reactions
  NEBNext® ARTIC SARS-CoV-2 Primer Mixes E7871S -20 1 x 24 reactions
  NEBNext® ARTIC SARS-CoV-2 FS Library Prep Kit (Illumina®) E7658-2 -20 1 x 96 reactions
  NEBNext® Sample Purification Beads E7659L 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 depth
  • No requirement for amplicon normalization prior to library preparation
  • Library inserts in the 150 bp range
  • Compatible with 2 x 75 Illumina sequencing
  • Optional use control human primers provided
  • Includes NEBNext Sample Purification Beads (SPRIselect®)
  • Adaptors and primers available separately; For a 384 unique dual index format, please contact us
An exception occurred during the operation, making the result invalid. Check InnerException for exception details.

Companion Products
References
  • Josh Quick nCoV-2019 sequencing protocol v2 (GunIt). protocols.io.
FAQs
Citations
Additional Citations
  • Mallm J-P, et al. (2021) Local emergence and decline of a SARS-CoV-2 variant with mutations L452R and N501Y in the spike protein. medRxiv DOI: 10.1101/2021.04.27.21254849
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 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.

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