Whole exome seq of mus musculus breast tissue; control for 3KO
Tumor cells isolated from mice were cultured in DMEM/F12 with 10% bovine growth serum. One 10 cm dish was used for each cell line to isolate genomic DNA using the DNEasy kit with an RNAse digestion step (Qiagen). For controls, breast tissue from adult virgin females was used to make genomic DNA using the same kit. DNA quantity and quality were verified using a Nanodrop. Whole exome sequencing libraries were prepared using the Agilent SureSelect XT library preparation kit according to the manufacturerÕs recommendations. DNA was sheared using a Covaris LE220. End-repaired, adenylated DNA fragments were ligated to Illumina sequencing adapters and amplified by PCR. Exome capture was performed using the Agilent SureSelect Mouse All Exon (50mb) capture probe set; captured exome libraries were enriched by PCR. Final libraries were quantified using the KAPA Library Quantification Kit (KAPA Biosystems), Qubit Fluorometer (Life Technologies) and Agilent 2100 BioAnalyzer, and were sequenced on an Illumina HiSeq2500 sequencer using 2x125 bp cycles.
3KO Control