Twist Oncology CGP Panel, Trinity Freestyle™ Workflow, 2x150 sequencing (Trinity Freestyle Sequencing Kit)

Methods

The EF 2.0 Library Preparation Kit (Twist Bioscience) was used to prepare libraries and perform capture via the Trinity Freestyle workflow according to the Twist Fast Hyb & Trinity Freestyle Run Setup Quick Guide. 

Twenty four libraries were prepared from 50 ng of human genomic DNA (NA12878), according to the EF 2.0 Library Preparation protocol and using the Twist Adapter System – Trinity Freestyle (#126889). A total of 12 libraries were pooled equally (500 ng per sample) per 1-hour fast hybridization capture reaction with 4 µL of the Twist Oncology Comprehensive Genomic Profiling (CGP) panel and Twist Trinity Freestyle Fast Hybridization Kit reagents (#127554). Two total hybridization reactions were prepared. 

Following capture, the hybridization reactions were diluted and pooled and the entire pool (400 µL) was sequenced on a single Trinity flow cell with a Trinity Freestyle 2x150 Sequencing Kit (#860-00046). Bases2Fastq was used to generate FASTQ files from the sequencing data. The sequencing run generated 950 million PF reads (286.57 Gb) with 94% Q30.