GSM7774802: AlphaToxin_U2OS_clone20; Homo sapiens; OTHER
genomic DNA (gDNA) was isolated from 5-10 X 106 cells 4 mg of gDNA were digested with BfaI or CviQI (New England Biolabs, Ipswich MA, USA) and purified (QIAquick PCR purification kits, Qiagen, Hilden, Germany) then ligated with barcoded P7-side adaptors using T4 DNA Ligase (New England BioLabs). Ligated gDNA fragments were then combined and amplified by nested PCR. The first round of PCR used primers corresponding to transposon sequence (KS15) and the P7-side adaptor (LP1). A second round of PCR (primers P2A and LP22) added the full Illumina P5 and P7 sequences. Sequencing was performed by the BRI Genomics platform using a HiScan or HiSeq platform (Illumina, San Diego CA, USA) with a custom sequencing primer (Rd1SeqP.ipdC) to generate 50 or 100 bp single end reads. described in Bruchez et al 2020