GSM5134216: GSCLC_S2; Mus musculus; Bisulfite-Seq
The sample preparations from GSCLCs were performed essentially as described previously (Ishikura et al., 2016) (see Fluorescence-activated Cell Sorting (FACS)).  15,000 cells of GSCLCs (GSCLC_S1, GSCLC_S2, GSCLC_S3, GSCLC_S4) were sorted and the genomic DNA was extracted with NucleoSpin ® Tissue XS (MACHEREY-NAGEL, U0901).  For the bisulfite treatment, the whole elute of the genomic DNA extraction containing 0.45 ng of unmethylated λ phage DNA was treated with an EZ DNA Methylation‐Gold Kit (ZYMO RESEARCH,  D5005) according to the manufacturer's instructions. Sequence libraries were generated with a post-bisulfite adaptor tagging method (http://www.crest-ihec.jp/english/epigenome/index.html; Miura et al., 2012; Shirane et al., 2016).  Some reagents were replaced as follows: Phusion Hot Start High‐Fidelity DNA Polymerase (Thermo Fisher Scientific, F540S) was replaced with Phusion Hot Start II DNA Polymerase (Thermo Fisher Scientific, F549S); Agencourt AMPure XP was replaced with an AxyPrep MAG PCR Clean‐Up Kit (Corning, MAG‐PCR‐CL‐250).  The sequencing was performed on an Illumina HiSeq 2500 platform with the TruSeq SR Cluster Kit ‐cBot‐HS and TruSeq SBS Kit ‐HS (Illumina). library strategy: Post-bisulfite adaptor-tagging (PBAT)