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 of human genes where baits have been carefully designed and optimized to provide complete coverage of the full coding (exon) 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.
Select your genes from our extensive library!
Target enrichment, coupled with next generation sequencing (NGS), enables high-throughput, deep sequencing of genomic regions of interest. NEBNext Direct is a novel, hybridization-based capture method offering significant advantages over traditional in-solution hybridization and multiplex PCR protocols.
In the NEBNext Direct target enrichment approach (Figure 1), enzymatically nicked or Covaris® sheared DNA is hybridized to biotinylated oligonucleotide baits that capture both strands of the target DNA and define the 3´ ends of the regions of interest. After hybridization, the bait-target hybrids are bound to streptavidin beads and any 3´ off-target sequence is removed enzymatically. This combination of hybridization with enzymatic removal of 3´ off-target sequence enables greater sequencing specificity relative to conventional hybridization-based enrichment methods. The trimmed targets are then converted into Illumina-compatible libraries that include a 12 bp unique molecular identifier (UMI) in the Illumina i5 index location and an 8 bp sample barcode in the Illumina i7 index location. The NEBNext Direct enrichment method can be performed within one to two days and is compatible with most automated liquid handling instruments.
The NEBNext Direct Custom Ready Panels are designed to enrich for the complete exonic content of genes selected from the Custom Ready portfolio for next generation sequencing on the Illumina platform. This kit contains the oligonucleotides, beads, enzymes and buffers required to convert the desired fragments into a sequence-ready library. Custom Ready Panels are designed for PE150 sequencing.
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 Direct® Index Primer Mix D01||-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||10 µM|
|NEBNext Direct® Index Primer Mix D05||10 µM|
|NEBNext Direct® Index Primer Mix D07||10 µM|
|NEBNext Direct® Index Primer Mix D08||10 µM|
|NEBNext Direct® Index Primer Mix D09||10 µM|
|NEBNext Direct® Index Primer Mix D10||10 µM|
|NEBNext Direct® Index Primer Mix D11||10 µM|
|NEBNext Direct® Index Primer Mix D12||10 µM|
|NEBNext Direct® Index Primer Mix D13||10 µM|
|NEBNext Direct® Index Primer Mix D14||10 µM|
|NEBNext Direct® Index Primer Mix D15||10 µM|
|NEBNext Direct® Index Primer Mix D16||10 µM|
|NEBNext Direct® Index Primer Mix D17||10 µM|
|NEBNext Direct® Index Primer Mix D18||10 µM|
|NEBNext Direct® Index Primer Mix D19||10 µM|
|NEBNext Direct® Index Primer Mix D20||10 µM|
|NEBNext Direct® Index Primer Mix D21||10 µM|
|NEBNext Direct® Index Primer Mix D22||10 µM|
|NEBNext Direct® Index Primer Mix D23||10 µM|
|NEBNext Direct® Index Primer Mix D24||-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|
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