title: GSM2584468: PFV intasome in vitro control; Homo sapiens; OTHER
library_construction_protocol: Genomic DNA extracted non-infected HT1080 cells using a GenElute kit (Sigma-Aldrich) was incubated with PFV intasome assembled with recombinant integrase and synthetic U5 donor DNA. The reaction products were deproteinized and repaired with Phi29 DNA polymerase (New England Biolabs). To amplify integration sites, DNA was digested with BfaI/BanII/CviQI and ligated to a linker made by annealing oligonucleotides 5´- TAGTCCCTTAAGCGGAG (with 3´ end blocked by an amino group modifier) and 5´-GTAATACGACTCACTATAGGGCCTCCGCTTAAGGGAC. PFV integration sites were amplified using primers 5´- CAAGCAGAAGACGGCATACGAGATCGGTCTCGGCATTCCTG CTGAACCGCTCTTCCGATCTGTAATACGACTCACTATAGGGC and 5´-GTGTGAA CTACACTTATCTTAAATGATG followed by a nested PCR with primers 5´- CAAGCAGAAGACGGCATACGAGATCGGTCTCGGCATTCCTGCTGAACCGCTCTT CCGATCTGTAATACGACTCACTATAGGGC 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: GSM2584468