Targeted Sequencing, xGen™ Hybridization Capture with xGen™ Pan-Cancer Hyb Panel (Trinity™ Sequencing Kit)

The Trinity sequencing kit was used to perform targeted sequencing on reference genomic DNA (gDNA).

Methods

Libraries were prepared using the xGen DNA Library Prep EZ UNI Kit (#10009822) with the xGen Stubby Adapter UDI primers for Element (#10017037). Capture for Trinity sequencing was performed according to the user guide with the xGen Pan-Cancer Hyb Panel (#1056205) probes.

Libraries were prepared from 100 ng of NA12878 gDNA, with 7 PCR cycles performed for amplification and indexing of the replicates. An additional 5 cycles of end-polishing PCR were performed on these libraries individually. A total of 24 libraries were pooled equally by mass (1000 ng per sample) in a single overnight hybridization capture reaction.

Following capture, the libraries were sequenced using the Trinity 2x150 Sequencing Kit (#860-00020). Bases2Fastq was used to generate FASTQ files from the sequencing data.