title: DNA-Seq of homo sapiens: lymphoma patients Fresh tissue lymphnode
design: 500ng of genomic DNA from each sample were sheared, and the size of 250-300bp fragmented DNA were purified by Beckman Ampure Bead. Libraries were constructed using ACCEL-NGS 2S DNA library kit from Swift Biosciences. Library preparation starts with Repair I, II and Ligation I, II. The indexed adapter is added during the Ligation step I to allow pooling of multiple libraries for hybridization and sequencing. PCR amplification was performed for 4 cycles and the concentration of the pre-libraries were quantified by Qubit double strand BR assay. IDT targeted probe (258 genes) captures were hybridized as pools of 8 samples using 150ng of library input, respectively. All hybridization and post captured washes were performed according to IDT protocol. Post PCR was performed using the Kapa HiFi DNA polymerase for 11-12 cycles. After the libraries were constructed, the Agilent TapeStation 4200 can be used to check the fragment size and concentration with the D1000 screen tape. A size distribution is between 380-430 bp and the final concentration is over than 5ng/l. Moreover, each pooling samples were quantified using KAPA Library Quantification Kit. Sequencing was performed on the Illumina NovaSeq6000 platform with 2x150 read lengths by following the manufacturer's protocols.