NEBNext Direct® BRCA1/BRCA2 Panel

Target with precision. 

NEBNext Direct employs a unique hybridization-based enrichment workflow that hybridizes baits directly to genomic DNA, without the need for upfront library preparation. The BRCA1/BRCA2 panel demonstrates extremely high specificity and unmatched coverage uniformity across a wide range of DNA inputs, allowing highly sensitive calling of germline and somatic variants while maximizing sequencer efficiency. 

  • Generate full (100%) coverage of all protein coding regions in BRCA1 and BRCA2 genes
  • Obtain highly uniform sequencing across exon targets- 100% of base pairs in panel have coverage greater than 20% of the mean target coverage
  • Maximize efficiency with a 1-day workflow that combines highly specific enrichment with library preparation
  • Produce high depths of target coverage across a wide range of DNA input amounts for germline and somatic variant calling

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  • Product Information

    BRCA1_Sample_2column_Ban_0717rev

    The NEBNext Direct BRCA1/BRCA2 Panel enriches the complete exon content of BRCA1 and BRCA2 genes for next-generation se­quencing analysis. NEBNext Direct employs a unique hybridization-based enrichment workflow that hybridizes baits directly to genomic DNA, without the need for upfront library preparation. The BRCA1/BRCA2 panel demonstrates extremely high specificity and unmatched coverage uniformity across a wide range of DNA inputs, allowing highly sensitive calling of germline and somatic variants while maximizing sequencer efficiency.

    Figure 1. NEBNext Direct Workflow.
    fast hybridization workflow

    The NEBNext Direct BRCA1/BRCA2 Panel delivers highly efficient enrichment of BRCA1 and BRCA2 coding regions with a high percentage of reads mapping to targets
    NEBNext Direct BRCA1/BRCA2 Panel
    This histogram shows the percent of reads aligned to the human genome, and the percent of reads mapped to the targets included in the BRCA1/BRCA2 Panel across different input DNA amounts. 10 ng, 100 ng and 1 µg of purified genomic DNA was enriched using the NEBNext Direct BRCA1/BRCA2 Panel. Sequencing reads were generated on an llumina® MiSeq® with 2x75 bp reads, 8 bp Sample ID and 12 bp unique molecular identifier. Sequencing read alignments were performed with BWA-MEM, and PCR duplicates were filtered using the UMIs.


    The NEBNext Direct BRCA1/BRCA2 Panel produces highly uniform coverage across target regions, maximizing sequencer efficiency
    specificity of BRCA testing histogram
    This histogram shows the normalized mean coverage across each of the 49 exon targets of the BRCA1 and BRCA2 genes. 50 ng of DNA was enriched using the 17 kb NEBNext Direct BRCA1/BRCA2 Panel. Sequencing reads were generated on an Illumina MiSeq with 2x75 bp reads, an 8 bp sample ID and 12 bp unique molecular identifier. Sequencing read alignments were performed with BWA-MEM, and PCR duplicates were filtered using the UMIs.


    The NEBNext Direct BRCA1/BRCA2 Panel can be used to provide depths of sequencing compatible with both germline and somatic variant calling
    BRCA Converge Uniformity Histogram
    This figure demonstrates de-duplicated coverage of BRCA1 and BRCA2 genes as a function of sequencing depth across a range of input DNA amounts. Purified genomic DNA was enriched using the NEBNext Direct BRCA1/BRCA2 Panel. Sequencing reads were generated on an Illumina MiSeq with 2x75 bp reads, the 8 bp sample ID and 12 bp unique molecular identifier. Sequencing read alignments were performed with BWA-MEM, and PCR duplicates were filtered using the UMIs.


    The NEBNext Direct BRCA1/BRCA2 Panel produces even depths of coverage across target regions
    coverage of BRCA1 and BRCA2 genes graph

    Integrative Genome Viewer (IGV) plot showing the read coverage across exon 1 of the BRCA1 gene. 50 ng of DNA was enriched using the 17 kb NEBNext Direct BRCA1/BRCA2 Panel. Sequencing reads were generated on an Illumina MiSeq with 2x75 bp reads, an 8 bp sample ID, and 12 bp unique molecular identifier. Sequencing read alignments were performed with BWA-MEM, and PCR duplicates were filtered using the UMIs.

    NEBNext Direct® BRCA1/BRCA2 Resources:

     

    Kit Components

    The following reagents are supplied with this product:

    Store at (°C) Concentration
    NEBNext Sample Purification Beads
    NEBNext Direct BRCA1/BRCA2 Baits -20
    NEBNext Direct® Bead Prep Buffer 4
    NEBNext Direct® Hybridization Buffer 4
    NEBNext Direct® dA-Tailing Buffer 4
    NEBNext Direct® Adaptor Ligation Buffer 4
    NEBNext Direct® Cleaving Buffer 4
    NEBNext Direct® Hybridization Wash (HW) 4
    NEBNext Direct® Bead Wash 1 25
    NEBNext Direct® Bead Wash 2 4
    NEBNext Direct® 3´ Adaptor -20
    NEBNext Direct® 5´ UMI Adaptor -20
    NEBNext Direct® dA-Tailing Enzyme -20
    NEBNext Direct® Ligase -20
    NEBNext Direct® 5´ Blunting Enzyme Mix -20
    NEBNext Direct® Cleaving Enzyme Mix -20
    NEBNext Direct® Q5 Master Mix -20
    NEBNext Direct® FFPE Phosphorylation Enzyme -20
    NEBNext Direct® Streptavidin Beads 4
    NEBNext Direct® Hybridization Additive -20
    NEBNext Direct® 5´ Blunting Buffer 4
    NEBNext Direct® 3´ Blunting Enzyme Mix -20
    Product Categories:
    Targeted Enrichment
    Applications:
    dna-library-preparation,
    Sample Preparation for Next Generation Sequencing,
    illumina-library-preparation
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