title: GSM5252650: Day 12 rep 1; Homo sapiens; OTHER
library_construction_protocol: CRISPR: Genomic DNA (gDNA) was isolated using the QIAamp DNA Blood Maxi Kit (Qiagen, 51192) as per the manufacturer's instructions. The gDNA concentrations were quanti-tated by Nanodrop. For PCR amplification, 252 μg of DNA was amplified per sample using the MyTaq Red Mix (Bioline, 25043). Reactions were set up in 384-well plates, using 200 ng of gDNA in 12 μL of reaction mix per well. Primers used for gDNA ampli-fication were Fwd 5' tgtggaaaggacgaaacacc 3' and Rev 5' cggactagccttattttaacttgc 3'. PCR cycling condition is as follows: an initial 2 minutes at 94 °C; followed by 15 seconds at 94 °C, 15 seconds at 56 °C, 30 seconds at 72 °C, for 30 cycles; and a fi-nal 5 minutes extension at 72 °C. P5/P7 primers were synthesized at Integrated DNA Technologies (IDT). PCR products were purified using the Agencourt AMPure XP SPRI beads (Beckman Coulter, A63880) according to manufacturer's instructions. Samples were sequenced on a NovaSeq S4 2X150 platform (Illumina).
attributes.tag: GEO Accession
attributes.value: GSM5252650