title: GSM7256452: Batch4_Ctrl.CBF6_S6; Homo sapiens; OTHER
library_construction_protocol: All blood samples were stored at -80°C and then thawed on ice prior to use. Plasma was obtained by centrifuging whole blood at 1,350 × g for 12 minutes. Following centrifugation, plasma was removed from the pellet via pipette and recentrifuged at 1,350 × g for 12 minutes and then at 13,500 × g for 5 minutes to ensure full removal of pellet. cfDNA was isolated from 1 to 2 mL of plasma using the QIAamp Circulating Nucleic Acid Kit (Qiagen) following manufacturer's instructions. Final elution was done using 25 μL of IDTE pH 8.0 buffer and purified DNA was stored at -80°C until use. In summary, sequencing adapters were ligated using the KAPA Hyper Prep Kit (KAPA Biosciences). cfDNA subsequently underwent T4 β-glucosyltransferase enzymatic modification with UDP-N3-glucose, followed by chemical modification with biotin-PEG4-dibenzocyclooctyne (DBCO). Biotin-labeled cfDNA fragments were then pulled down using streptavidin M270 Dynabeads (Invitrogen). Resulting pulled down DNA was then PCR amplified to construct sequencing libraries. Libraries were purified using AMPure XP beads (Beckman Coulter) following a standard purification protocol.