GSM4983817: Mouse_ear; Mus musculus; OTHER
Mouse tissues were digested with proteinase K at 50°C overnight and the genomic DNA was precipitated with ethanol in the presence of sodium chloride. One microgram of genomic DNA was fractionated by gel electrophoresis to remove the majority of the episomal viral genomic DNA. The host DNA was recovered and sheared into 300-500 bp long fragments using a Covaris M220 Focused-ultrasonicator (Covaris). The fragmented DNA was end repaired and dA-tailed using the NEBNext Ultra Ligation kit (NEB, #E7445) according to the manufacturer's instructions. A partially double stranded linker (5'-GTAATACGACTCACTATAGCGCTGCGCTTAAGCGACT-3', 5'–PO4-GTCGCTTAAGCGCAG-3'-C6) with a 3' T overhang was ligated to the genomic DNA fragments using the NEB Quick Ligation Kit (NEB, #M220). Virus specific primers (Pr2976/Pr3277/Pr3742) were used to selectively amplify the integration junctions. Two rounds of 20 cycles PCR were conducted to enrich integration junctions in the sequencing libraries using Phusion High-Fidelity PCR Kit (NEB, #E0553).