GSM8140414: WGBS-HG00621-Rep2; Homo sapiens; Bisulfite-Seq
Qiagen AllPrep DNA/RNA kit 200ng of gDNA, including 0.2% Unmethylated Lambda DNA (Promega, Cat# D1521) were fragmented in a final volume of 50uL using the Covaris LE220 targeting ~350bp inserts. A 1.5x AMPure clean-up was performed post fragmentation on the EpMotion 5075 (Eppendorf) resulting in a final volume of 20uL. The fragmented gDNA was bisulfite converted with the EZ-96 DNA Methylation-Gold Mag Prep Kit (Zymo Research, Cat # D5043) according to the manufacturer's recommendations on the EpMotion 5075. The bisulfite converted DNA (bsDNA) was quantitated on the Qubit Flex using the Qubit ssDNA Assay Kit (Thermo Fisher Scientific, Cat #Q10212). Automated whole genome bisulfite (WGBS) libraries were constructed with ~100ng of bsDNA using the Accel-NGS Methyl-Seq DNA Library Kit (Swift BioSciences, Cat # 30096) and unique dual indexes (UDI) (Swift, X9096) on the EpMotion 5075. Eight PCR cycles will be performed during the indexing PCR step, followed by a final 0.85x AMPure cleanup.