title: GSM9055194: 0.5 mM DFMO treated for 14 days; Homo sapiens; OTHER
library_construction_protocol: Genomic DNA (gDNA) was extracted from cell pellets of 20 x 106 cells using the Blood genomicPrep Mini Spin Kit (Cytiva # 28904264) or 60 × 106 cells using the QIAamp DNA Blood Maxiprep Kit (Qiagen # 51192) according to manufacturer's instructions with minor modifications: Cytiva and QIAGEN Protease were replaced with a 10 mg/mL solution of ProteinaseK (MilliporeSigma # 3115879001) in water; cells were lysed overnight; centrifugation steps after Qiagen Buffer AW1 and AW2 were performed for 2 minutes and 5 minutes, respectively;  2 x 30 µL (Cytiva) or 1 mL (Qiagen) of water preheated to 70 °C was used to elute gDNA (1- or 5-minute incubation), followed by centrifugation for 1 or 5 minutes (Cytiva or Qiagen extractions, respectively). gDNA was quantified using the Qubit dsDNA HS Assay kit (Thermo Fisher Scientific #Q32851). All PCR reactions were performed in 50 µL reactions using ExTaq Polymerase (Takara Bio #RR001B). For all samples, 1, 3, or 6 µg of gDNA was initially amplified for 28 cycles in 50 µL test PCR reactions. Subsequently, an additional 50 µL reactions were performed using 6 µg per reaction (140 µg gDNA). Reactions were pooled and 100 µL of each sample was purified using HighPrep PCR beads (MagBio Genomics #AC-60005), eluted with 20 µL water, and quantified using the Qubit dsDNA HS Assay kit before sequencing for 26 cycles on an Illumina NovaSeq.