Targeted sequencing in mouse lung tumors. Mus musculus

We applied Cas9-mediated targeted mutation in mouse lung tissues, where tumors might develop. The genomic DNA was isolated from either freshly dissected lung tumours or lobes using the QIAamp DNA mini kit (Qiagen). For each target gene or potential off-target sites, a genomic region containing the target sequence was PCR-amplified using specific primers and gel purified, followed by library construction and high-throughput sequencing analysis (Illumina Hiseq 2500).