GSM9313419: HIV-1, Naked target DNA (fragmented by sonication), rep2; Saccharomyces cerevisiae; OTHER
organic extraction Chromatinized or naked yeast DNA was incubated with purified retroviral intasomes. Chromatinized reaction products were fragmented using Fenton reaction in the presence of copper (II) cloride, mercaptopropionic acid, and hydrogen peroxide. DNA was then deproteinized,  repaired with phi29 DNA polymerase and processed for dA tailing. End-prepped DNA was ligated to a linker (obtained by annealing 5'-phosphorylated and 3'-blocked 5'-GTCCCTT AAGCGGAG with 5'- GTAATAC GACTCAC TATAGGGCC TCCGCTT AAGGGACT). The products,  PCR amplified using primers specific to the linker and viral DNA ends, were sequenced using Nanopore technology.