title: GSM7669335: BCAR-T Donor 3; Homo sapiens; Bisulfite-Seq
library_name: GSM7669335
library_construction_protocol: Genomic DNA from samples was isolated with QIAGEN DNeasy Blood & Tissue kit following the corresponding manufacturer's protocols. The isolated genomic DNA was sonicated by Covaris system. Size selection by Ampure XP beads (Beckman-Coulter) was performed to enrich DNA fragments of around 250 bp in average sizes before library construction. In brief, 0.7 volume beads were added to the fragmented genomic DNA. After incubation, the supernatant was transferred to a new tube and an additional 1.3 volume beads were added (“0.5×–2× bead size selection”). After washing two times with 80% ethanol, genomic DNA was eluted with 40 ul EB buffer. Subsequently, 200ng genomic DNA was used for DNA library preparation with NEBNext Ultra II DNA library prep Kit (Cat#E7645). DNA library preparation has the following steps: DNA fragments end prep, adaptor ligation, size selection or purification of Adaptor-ligated DNA with Ampure XP beads, bisulfite conversion by Epitect Fast DNA Bisulfite Kits (QIAGEN, Cat#59826), PCR enrichment of Adaptor-ligated DNA, and purification of PCR product with 0.9x Ampure XP beads. The average size of resulting DNA fragments was around 480bp, validated by Tapestation. The libraries were then subjected to sequencing on Illumina NovaSeq sequencer with 2x150bp configuration (Azenta).