title: GSM8735975: oncogene negative tumor, Pten guide1 site1, 9 weeks, Sample_ID LA75_61; Mus musculus; OTHER
library_construction_protocol: Bulk tumor-bearing tissues were homogenized in 6 mL of cell lysis buffer (Puregene, 158063, Qiagen) using a FastPrep-24 5G tissue homogenizer (116005500, MP Biomedicals) and digested overnight with proteinase K (AM2544, Life Technologies). Benchmark control cell lines containing unique barcodes (50,000 or 100,000 cells each) were added to each lung sample prior to lysis to enable the calculation of the absolute number of neoplastic cells in each tumor. Q5 High-Fidelity 2x Master Mix (M0494X, New England Biolabs) was used to amplify the U6-BC-crRNA region from 32 μg of genomic DNA in a total reaction volume of 800 μL per sample using unique dual-indexed primers.