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

NEBNext ARTIC SARS-CoV-2 kits are also available for larger (~400bp) insert libraries for Illumina sequencing (NEB #E7650), and for Oxford Nanopore Technologies® sequencing (NEB #E7660). 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. 

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 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 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. 

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. 

  • 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
  • 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

 

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

 
Catalog # Concentration Size
E7658S Not Applicable 24 reactions
E7658L Not Applicable 96 reactions
To find out how to order this product from your current location, click the button below:
International Customer & Order Support
Please enter a quantity for at least one size

Want to try this product? Request a sample Need a custom/large volume order? Contact Us

Bulk packaging may also be available and requested for large recurring orders.
Learn More

  • Product Information
    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. The V3 ARTIC primers have been balanced, using methodology developed at NEB based on empirical data from sequencing, to provide uniform amplicon coverage across the viral genome. 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 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 sequencing. 

    Figure 1: Improved genome coverage with the NEBNext ARTIC SARS-CoV-2 FS Library Prep Kit (Illumina)



    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, with or without NEBNext ARTIC Human Control Primer Pairs 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 with seqtk and aligned to SARS-CoV-2 reference genome (NCBI, NC_045512) with Bowtie2.



    Figure 2: Improved coverage depth per base with NEBNext ARTIC SARS-CoV-2 FS Library Prep Kit (Illumina)  



    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 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 FS Library Prep Kit (Illumina) and sequenced on a MiSeq instrument (2x250 bp). Coverage depth per base was determined, reads were down-sampled with seqtk and aligned to SARS-CoV-2 reference genome (NCBI, NC_045512) with Bowtie2.
     


    Figure 3: Including human control primers does not adversely affect genome coverage 



    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 NEBNext balanced ARTIC SARS-CoV-2 primer pools, with or without NEBNext ARTIC Human Control Primer Pairs. Libraries were constructed using the NEBNext ARTIC SARS-CoV-2 FS Library Prep Kit (Illumina) and sequenced on a MiSeq instrument (2x75 bp). The fraction of the genome covered at each depth was determined for a range of inputs and reads down-sampled to 10,000, 100,000, 500,000 and 1,000,000.


    Figure 4: Improved genome coverage with the NEBNext ARTIC SARS-CoV-2 FS Library Prep Kit (Illumina)



    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 QIAseq SARS-CoV-2 Primer Panel or the NEBNext balanced ARTIC SARS-CoV-2 primer pools. Libraries were constructed using the QIAseqFX kit or 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 with seqtk and aligned to SARS-CoV-2 reference genome (NCBI, NC_045512) with Bowtie2.


    Figure 5: Improved genome coverage with the NEBNext ARTIC SARS-CoV-2 FS Library Prep Kit (Illumina) 



    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 QIAseq SARS-CoV-2 Primer Panel or the NEBNext balanced ARTIC SARS-CoV-2 primer pools. Libraries were constructed using the QIAseqFX kit or the NEBNext ARTIC SARS-CoV-2 FS Library Prep Kit (Illumina) and sequenced on a MiSeq instrument (2x75 bp).The fraction of the genome covered at each depth was determined for a range of inputs and reads down-sampled to 10,000, 100,000, 500,000 and 1,000,000.


    Figure 6: NEBNext ARTIC SARS-CoV-2 FS DNA (Illumina) Workflow 



    Kit Components

    The following reagents are supplied with this product:

    NEB # Component Name Component # Stored at (°C) Amount Concentration
    • E7658S     Multi-temperature    
    • E7658L     Multi-temperature    
    Product Categories:
    NGS Sample Prep & Target Enrichment Products
    Applications:
    NEBNext® ARTIC products for SARS-CoV-2 sequencing,
    RNA For Illumina,
    RNA Library Preparation,
    RNA-seq,
    NGS Sample Prep & Target Enrichment,
    Illumina Library Preparation
    • Advantages and Features
    • Properties & Usage
    • Related Products
    • References
  • Protocols, Manuals & Usage
  • Tools & Resources
  • FAQs & Troubleshooting
  • Citations & Technical Literature
    • Citations
  • Quality, Safety & Legal
Loading Spinner