NEBNext Direct® Custom Ready Panels

Use our NEBNext Direct® Custom Ready Panel Request Form to order. 
 
 

Flexible Precision.

Employing the unique NEBNext Direct hybridization-based enrichment method, NEBNext Direct Custom Ready Panels allow rapid customization of targeted gene panels for Illumina® sequencing. Select from a list for which baits have been carefully designed and optimized to give complete coverage of the full coding regions. High quality panels can be designed by you and rapidly delivered, from any combination of genes. NEBNext Direct Custom Ready Panels provide the content you want with the performance you need.

  • Choose from a single gene to hundreds of genes. View Ordering Guide.  
  • Experience unmatched specificity and coverage uniformity
  • Eliminate synthesis and optimization steps, for a faster turnaround
  • Improve sensitivity with our Unique Molecule Index (UMI)
  • Generate results in one day with our automation-friendly workflow

Ordering Information

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E6631

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  • Product Information

    Select your genes from our extensive library! 

    Each NEBNext Direct Custom Ready Panel is designed to enrich for the complete exonic content of genes selected from the Custom Ready portfolio 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. NEBNext Direct Custom Ready Panels provide extremely high specificity and unmatched coverage uniformity across a wide range of DNA inputs, enabling highly sensitive calling of germline and somatic variants while maximizing sequencer efficiency.

    Figure 1: NEBNext Direct Workflow


    Fig1_NEBNext_Direct_CR_Workflow

     
    Figure 2: NEBNext Direct Custom Ready Panels demonstrate optimum performance across a wide range of panel sizes.


    Fig4_NEBNextDirectCR_Performance

    Key target enrichment metrics demonstrate consistent performance across a range of panel sizes. 100 ng of DNA was tested against panels of 1, 10, 25, 50 and 100 genes, and sequenced using Illumina paired-end 150 bp sequencing. Larger panels included all genes present in smaller panels.
    Figure 3: NEBNext Direct Custom Ready Panels demonstrate retention of target behavior across panel sizes.


    Fig3_NEBNextDirectCR_IGV

    IGV image of coverage profile for 4 BRAF exons included in panels of 1, 10, 25, 50 and 100 genes, demonstrate consistent target behavior with the addition of gene targets. 100 ng of DNA was used as input for NEBNext enrichment using the 5 panels, including the BRAF gene. Libraries were sequenced on an Illumina 2 x 150 basepair sequencing.
    Figure 4: Sensitivity in detection of variants across panel size and DNA input amount.


    Fig2_NEBNextDirectCR_VariantDetection

    24 HapMap samples were blended to create a range of variant allele frequencies (VAF) down to 2%. 25, 50, 100, 200 and 500 ng of this blended DNA was enriched using NEBNext Direct Custom Ready Panels of 1, 10, 25, 50, and 100 genes. Larger panels were inclusive of the genes in smaller panels. Resulting libraries were sequenced using 2 x 150 bp Illumina sequencing and variants were called using Mutect and Vardict variant calling algorithms.
    Figure 5: NEBNext Direct Nicking Enzyme produces consistent fragmentation across DNA input quantities.


    Fig5_NEBNextDirectCR_NickingOverlay

    10 ng, 100 ng, and 1 µg of Control DNA was used as input for fragmentation using the NEBNext Direct Nicking Enzyme. Representative electropherograms show fragment sizes compatible with NEBNext Direct enrichment across DNA input amounts.
    Figure 6: NEBNext Direct Nicking Enzyme performance vs. mechanical shearing


    Fig6_NEBNextDirectCR_NickingEnzyme

    100 ng of Control DNA was used as input to fragmentation using the NEBNext Direct Nicking Enzyme and Covaris® Shearing. Fragmented DNA was enriched using the 37 kb NEBNext Direct Cancer HotSpot Panel, and sequenced to a mean depth of 3 million reads using Illumina 2 x 75 bp sequencing.

    Reagents Supplied

    The following reagents are supplied with this product:

    Store at (°C) Concentration
    NEBNext Direct® Bead Prep Buffer 4
    NEBNext Direct® dA-Tailing Buffer 4
    NEBNext Direct® Adaptor Ligation Buffer 4
    NEBNext Direct® Cleaving Buffer 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 Index Primer Mix D01-D08 (E7020-E7027) -20 10 μM
    NEBNext Direct® Index Primer Mix D02 -20 10 μM
    NEBNext Direct® Index Primer Mix D03 -20 10 μM
    NEBNext Direct® Index Primer Mix D04 -20 10 μM
    NEBNext Direct® Index Primer Mix D05 -20 10 μM
    NEBNext Direct® Index Primer Mix D07 -20 10 μM
    NEBNext Direct® Index Primer Mix D08 -20 10 μM
    NEBNext Direct® Index Primer Mix D09 -20 10 μM
    NEBNext Direct® Index Primer Mix D10 -20 10 μM
    NEBNext Direct® Index Primer Mix D11 -20 10 μM
    NEBNext Direct® Index Primer Mix D12 -20 10 μM
    NEBNext Direct® Index Primer Mix D13 -20 10 μM
    NEBNext Direct® Index Primer Mix D14 -20 10 μM
    NEBNext Direct® Index Primer Mix D15 -20 10 μM
    NEBNext Direct® Index Primer Mix D16 -20 10 μM
    NEBNext Direct® Index Primer Mix D17 -20 10 μM
    NEBNext Direct® Index Primer Mix D18 -20 10 μM
    NEBNext Direct® Index Primer Mix D19 -20 10 μM
    NEBNext Direct® Index Primer Mix D20 -20 10 μM
    NEBNext Direct® Index Primer Mix D21 -20 10 μM
    NEBNext Direct® Index Primer Mix D22 -20 10 μM
    NEBNext Direct® Index Primer Mix D23 -20 10 μM
    NEBNext Index Primer Mix D01-D24 (E7020-E7043) -20
    NEBNext Direct® Index Primer Mix Plate (NEB #E7000X) -20
    NEBNext Sample Purification Beads 4
    NEBNext Direct® Streptavidin Beads 4
    NEBNext Direct® Hybridization Additive -20
    NEBNext Direct® 3´ Blunting Buffer 4
    NEBNext Direct® 5´ Blunting Buffer 4
    NEBNext Direct Hybridization Wash (HW) 4
    NEBNext Direct Hybridization Buffer 4
    NEBNext Direct® 3´ Blunting Enzyme Mix -20
    NEBNext Direct® DNA Nicking Buffer
    NEBNext Direct® DNA Nicking Enzyme
    NEBNext Direct® Stop Solution
    NEBNext Direct Custom Baits
    Product Categories:
    Targeted Enrichment,
    sample-preparation-for-next-generation-sequencing
    Applications:
    dna-library-preparation,
    Sample Preparation for Next Generation Sequencing
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