title: GSM9342215: RRBS hippocampus, WT, biological replicate 6; Mus musculus; Bisulfite-Seq
library_construction_protocol: Hippocampal tissue was lysed at 56°C overnight and genomic DNA was extracted using the QIAamp DNA Kit (Qiagen) according to the manufacturer's instructions. DNA was eluted in 50 µl elution buffer. DNA concentration was measured using the Quant-iT PicoGreen dsDNA Assay (Invitrogen) and DNA integrity was assessed with the Agilent TapeStation 4150 using the Genomic DNA ScreenTape assay. Reduced Representation Bisulfite Sequencing (RRBS) libraries were prepared from 100 ng genomic DNA using the Diagenode Premium RRBS Kit following the manufacturer's guidelines. The protocol included restriction digestion, size selection, bisulfite conversion, and PCR amplification. Methylated and unmethylated spike-in controls provided in the kit were used to estimate bisulfite conversion efficiency. Libraries were quantified using PicoGreen and fragment size distribution was assessed using the Agilent Bioanalyzer High Sensitivity DNA Kit. Libraries were denatured and diluted to 1.8 pM with 20% PhiX Control (Illumina) prior to sequencing.