title: GSM6205399: HTISA.ASLV.total.3; Homo sapiens; OTHER
library_construction_protocol: Total and GFP-negative cultures were harvested for genomic DNA isolation on the same day so that culture times after infection with GFP7 were the same. Genomic DNA was isolated using a salt-extraction method (Aljanabi and Martinez, 1997). HTISA libraries were prepared and sequenced from GFP-negative and Total populations ("B" culture genomic DNAs were combined to yield a GFP-negative "Pool" library and a Total "Pool" library, so that finally, we obtained one pooled and 4 separate GFP-negative samples and one pooled and 4 separate Total samples). We used a linear amplification–PCR protocol based on a previously published high-throughput, genome-wide, translocation sequencing (HTGTS) method (Frock et al., 2015). The adaptation of the protocol was as follows: Retroviral integration sites were linearly amplified from the vector 5' LTR using biotinylated Bio_L7a (HIV-derived vector integrations) or Bio_A7a (ASLV-derived vector integrations) primers. Nested PCR was performed with barcoded inner primer I5_bar75_L7a (HIV-derived vector integrations) or I5_bar75_A7a (ASLV-derived vector integrations). The blocking digestion of the nested PCR product was omitted. The resulting PCR product were gel purified and sequenced using a NextSeq500 (Illumina) sequencer.