HEK293T_targeted_deep_seq
Two-step PCR amplification to produce DNA fragments for each on-target and off-target site was used. In the first step, we used locus-specific primers bearing universal overhangs with complementary ends to the TruSeq adaptor sequences. In the second step, purified PCR products were amplified with a universal forward primer and an indexed reverse primer to reconstitute the TruSeq adaptors. Full-size products (~250bp in length) were purified. The purified library was deep sequenced using a paired-end 150bp MiSeq run.