title: GSM8986228: P8, 64 years, Colon; Homo sapiens; Bisulfite-Seq
library_construction_protocol: DNA extraction using the QIAmp DNA mini kit (Qiagen, Hilden, Germany) according to manufacturers instructions 300 ng of genomic DNA were bisulfite converted using the EpiTect Fast Bisulfite conversion kit (Qiagen, Hilden, Germany) according to manufacturers instructions. 3 µl of Bisulfite coverted DNA was subsequently used in first round of PCRs in 50 µl volume reactions for target enrichment. 0 % and 100 % methylation controls served as controls for assessing the reliability of methylation measurements. The samples were quantified using the Qubit fluorometer and diluted to a concentration of 0.2 ng/µl. In the subsequent PCRs, samples were barcoded using NEBNext Multiplex Oligos for Illumina (Dual Index Primers Set 2). The purified and quantified PCR pools were diluted to a concentration of 4 nM, and 3 µl of this dilution from each of the MIDs were combined into a single final pool for next-generation sequencing, using the NextSeq 2000 platform (Illumina, California, USA) and Reagent Kit P1 (300 cycles) cartridge (Illumina) according to the manufacturer's instructions.