GSM2803046: T3_d; Mus musculus; Bisulfite-Seq
3-4x105 sorted T0-T3-SCs cells were lysed and their genomic DNA extracted using the RecoverAll Kit for FFPE (Ambion, AM1975) following the manufacturer's guidelines (one animal per replicate, n=4). Genomic DNA samples were first concentrated with Genevac EZ-2 Evaporation System and diluted in water to reach 95ng in 26µl. Sequencing library pool was prepared with the Premium Reduced Representation Bisulfite Sequencing (RRBS) Kit (Diagenode, C02030033) following manufacturer's instructions, quantified with the Qubit dsDNA HS Assay Kit (Thermo Fisher Scientific, Q32854) and average size of DNA fragments was estimated with a 2100 Bioanalyzer instrument using Agilent High Sensitivity DNA Kit (5067-4626). Library pool was then denaturated in NaOH and diluted at 1.9pM with 20% of PhiX Control v3 (Illumina, FC-110-3001). To improve cluster density consistency, the diluted library pool was heat-denatured at 96°C for 2min, inverted twice to mix, and quickly moved to ice for 5min before loading onto the sequencer. The library pool was sequenced on a NextSeq 500 (Illumina) using the NextSeq 500/550 High Output v2 Kit, 75 cycles (Illumina, FC-404-2002). Sequencing was single-read, 76 cycles, index 1: 6 bases.
GEO
GSM2803046