title: Integration sites from MLV-infected wild-type HEK293T cells
design: Sowd et al. (PNAS. 2016) infected wild-type HEK293T cells with VSVg-pseudotyped MLV.  DNA was extracted from 2 to 5 million cells using the DNeasy Blood and Tissue Kit (Qiagen). Ten micrograms of DNA was digested with MseI and BglI, ligated to adapters, and amplified by ligation-mediated PCR as described by Matreyek et al. Retrovirology (2014).  A second round of nested PCR was performed to further amplify MLV LTR containing PCR fragments. Nested PCR primers contained unique linker sequences.  PCR products were sequenced using the Illumina MiSeq platform and single-end 150 bp next-generation sequencing.