title: GSM3035201: Bisulfite conversion and whole genome sequencing_veg; Paramecium tetraurelia; Bisulfite-Seq
library_construction_protocol: Cells were harvested and washed twice with 10mM Tris-HCl pH 7.4. The cell pellet was then incubated with the lysis buffer. Cells were incubated with 20μl of RNAse A (from GeneElute –Mammalian Genomic DNA Miniprep Kit Sigma-Aldrich) for five minutes at room temperature. Afterward, cells were incubated in three volumes of the lysis buffer (0.5M EDTA, 1% SDS, 1% N-lauryl sarcosine, 1 mg/ml of proteinase K) at 55°C for overnight. The cells were then incubated for 1hr with an equal volume of Phenol: Chloroform: IAA on a shaker with gentle shaking. After incubation, the cells were centrifuged at the maximum speed of the table top centrifuge for 10minutes. The supernatant was then transferred to a clean tube, and genomic DNA was precipitated using 800μl of isopropanol. Ovation Ultralow Methyl-Seq kit (Nugen) with ~0.5% Phi-X spiked in.
attributes.tag: GEO Accession
attributes.value: GSM3035201