title: GSM2584474: R540Q PFV (codon-optimized, 4-component) vector in HEK293T cells; Homo sapiens; OTHER
library_construction_protocol: DNA was extracted from infected cells five days post-infection using a Gentra Puregene kit (Qiagen). Cellular genomic DNA isolated 5 days post-infection was digested with BfaI/BanII/CviQI and ligated to a linker made by annealing oligonucleotides 5´- TACGATGCGGTAACTGC (with 3´ end blocked by an amino group modifier) and 5´-GTATCTCAACAAGCAGCTTGAGGCAGTTACCGCATCG. PFV integration sites were amplified using primers 5´- CAAGCAGAAGACGGCATACGAGATCGGTCTCGGCATTCCTGCTGAACCGCTCTTCCGATCTGTATCTCAACAAGCAGCTTGAG3' and 5´-GTGTGAACTACACTTATCTTAAATGATG followed by a nested PCR with primers 5´- CAAGCAGAAGACGGCATACGAGATCGGTCTCGGCATTCCTGCTGAACCGCTCTTCCGATCTGTATCTCAACAAGCAGCTTGAG and 5´- AATGATACGGCGACCACCGA GATCTACACTCTTTCCCTACACGACGCTCTTCCGATCTXXXXXCTTAAATGATGTAACTCCTTAGGATAATCAATATAC (the string of underlined nucleotides marked with “X” denotes the position of a 5-base index, which was unique for each integration site library analyzed). 
attributes.tag: GEO Accession
attributes.value: GSM2584474