Human Whole-Exome Sequencing, Trinity Freestyle™ Workflow, Twist Exome 2.0 + Comp Spike-in, 2x150 sequencing (Trinity Freestyle Sequencing Kit)

Dataset 1: 24 samples on AVITI

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. 

Libraries were prepared from 50 ng of human genomic DNA (NA12878), according to the EF 2.0 Library Preparation protocol. A total of 8 libraries were pooled equally by mass (500 ng per sample) per 1-hour fast hybridization capture reaction with 4 µL of the Twist Exome 2.0 panel + Comp Spike-in (#126855) and the Twist Trinity Freestyle Fast Hybridization Kit reagents (#127554). Three total hybridization reactions were prepared. 

Following capture, the hybridization reactions were pooled and 50 µL of the pool 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. 

Dataset 2: 32 samples on AVITI24 

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. 

Libraries were prepared from 50 ng of human genomic DNA (NA12878), according to the EF 2.0 Library Preparation protocol using the Twist Adapter System – Trinity Freestyle (#126889).  A total of 8 libraries were pooled equally by mass (500 ng per sample) per 1-hour fast hybridization capture reaction with 4 µL of the Twist Exome 2.0 panel + Comp Spike-in (#126855) and the Twist Trinity Freestyle Fast Hybridization Kit reagents (#127554).  Four total hybridization reactions were prepared. 

Following capture, the hybridization reactions were pooled and 60 µL of the pool was sequenced on a single Trinity flow cell with a Trinity Freestyle 2x150 Sequencing Kit (#860-00046) on AVITI24. Bases2Fastq was used to generate FASTQ files from the sequencing data.