GSM4432289: d6hPGCLC; Homo sapiens; Bisulfite-Seq
10,000-15,000 cells were lysed in the lysis buffer [0.1% of SDS and 1 µg/µL of Proteinase K in DNase-free water (GIBCO, 15230-160)] containing unmethylated lambda phage DNA (Promega, D1521) at 37°C for 60 min and then incubated at 98°C for 15 min to inactivate proteinase K. Assuming that a single cell bears 6 pg of genome DNA, we added the unmethylated lambda phage DNA equivalent to 0.5% of the input DNA. For the bisulfite treatment, the cell lysate was treated with EZ DNA Methylation-Gold Kit (Zymogen, D5005) according to the manufacturer's instruction. To construct sequence libraries, the bisulfite-treated DNA was subjected to the serial synthesis of the single-end adaptor-tagged complementary DNA fragments (http://www.crest-ihec.jp/english/epigenome/index.html).