GSM8425873: Day8CART_Biorep3; Homo sapiens; OTHER
To prepare for sequencing, 3.5x10^7 cells from baseline (Day 8) and chronically stimulated (Day 22) timepoints were washed and pelleted before storage at -20°C. Next, genomic deoxyribonucleic acid (gDNA) was isolated from each cell pellet with the Quick-DNA Midiprep Plus Kit (cat #D4075, Zymo Research, Irvine, CA, USA) according to the manufacturer's protocol. An ethanol precipitation protocol was performed to enhance the purity of the gDNA following isolation. Next, the gDNA was prepared for sequencing through library PCR. For the library PCR, the NEBNext® High-Fidelity 2X PCR Master Mix (cat #M0541S, New England BioLabs, Ipswich, MA, USA) was used with the primers and the cycling conditions that were developed and described in the referenced protocol paper for the GeCKO v2 library (https://doi.org/10.1038/nprot.2017.016).