title: GSM3671081: CRISPR-Cas9 screening experiment1 2C; Mus musculus; OTHER
library_construction_protocol: Genomic DNA of 2C-positive and negative cells were extracted through Qiagen DNeasy Blood & Tissue Kit (Cat. No. 69504). Around 1 million 2C-positive cells and 10 million 2C-negative cells were collected through FACS based on tdTomato reporter expression. Genomic DNA of 2C-positive and negative cells were extracted through Qiagen DNeasy Blood & Tissue Kit (Cat. No. 69504). gRNA sequences were then amplified using P5 primer (AATGATACGGCGACCACCGAGATCTACACTCTTT CCCTACACGACGCTCTTCCGATCTTTGTGGAAAGGACGAAACACCG) and P7 primer (5'CAAGCAGAAGACGGCATACGAGATNNNNNNNNGTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTCCAATTCCCACTCCTTTCAAGACCT; NNNNNNNN refers to 8bp index) through KAPA HiFi HotStart ReadyMix (Cat. No. KK2602). For each sample, 4 parallel PCR reactions were set up, and mixed products from 4 reactions were then purified through AMPure XP reagents (1.0x) and sequenced by an Illumina HiSeq 2500 Sequencer. Two screen replicates (independent Dux induction and FACS isolation) were performed.
attributes.tag: GEO Accession
attributes.value: GSM3671081