GSM7764393: RRBS, Ctrl, K107; Homo sapiens; Bisulfite-Seq
Genomic DNA extraction from whole blood. We prepared aliquots of 1 ml frozen whole blood sample from ALS and control patients and extracted 200 μL of it using the Macherey-Nagel™ NucleoSpin™ Tissue Kit according to the manufacturer's protocol. The premium RRBS kit of Diagenode was employed to carry out the RRBS protocol including NGS library preparation, as described (Veillard, AC., Datlinger, P., Laczik, M. et al. Diagenode® Premium RRBS technology: cost-effective DNA methylation mapping with superior coverage. Nat Methods 13, i–ii (2016). https://doi.org/10.1038/nmeth.f.391). Briefly, 100 ng of DNA was fragmented by the MspI restriction enzyme, which cleaves DNA without regard to the cytosine methylation state. Following adaptor ligation and size selection by Ampure beads, we pooled up to 6 samples together. These pooled samples underwent bisulfite treatment, purification, and PCR amplification, according to the manufacturer's recommendations. The final RRBS libraries were quantified using the Qubit dsDNA HS Assay from Life Technologies, and the library's size distribution profile was assessed using Agilent Bioanalyzer 2100 capillary electrophoresis.