WGS of mus musculus: IFNR knockout peripheral blood
Briefly, peripheral blood was collected in tubes with heparin sodium salt (Sigma, 2106) from the submandibular vein of the F0 male WT1xIFNR and two male C57BL/6NTac WT controls. Red blood cells were then lysed by ammonium chloride-potassium (ACK, (5)) then DNA isolated using the AllPrep DNA/RNA/Protein Mini Kit (Qiagen, Cat# 80004). DNA was quantified using the Qubit 3 Fluorometer according to manufacturers instructions (Invitrogen). Libraries were generated from 1 mg DNA using NEBNext DNA Library Prep Kit and indices were added to each sample. The genomic DNA is randomly fragmented to a size of 350bp by shearing, then DNA fragments were end polished, A-tailed, and ligated with the NEBNext adapter for Illumina sequencing, and further PCR enriched by P5 and indexed P7 oligos. The PCR products were purified (AMPure XP system) and resulted libraries were sequenced on an Illumina NovaSeq 6000 instrument