GSM8827307: Small intestinal crypt8, 22 weeks; Mus musculus; Bisulfite-Seq
Mouse Small-Intestine Crypts: Small-intestine crypts were isolated from two wild-type B6 mice (aged 22 and 28 weeks) as previously described (https://doi.org/10.1007/978-1-62703-125-7_19).DNA extraction was performed by resuspending each crypt in 20 µL of lysis buffer (20 mM Tris, 20 mM EDTA, 20 mM KCl, 0.3% Triton X in ddH₂O) with 80 µg of Proteinase K. Samples were incubated at 50°C for 3 hours, followed by heat inactivation at 85°C for 10 minutes. Whole-genome methylation libraries were prepared using the NEBNext Enzymatic Methyl-seq Kit (E7120). Human Hair Follicle Cells: Individual black or white hairs were plucked from a 53-year-old volunteer. The hairs were grouped into black and white categories and immediately subjected to DNA extraction with the QIAGEN DNeasy® Blood & Tissue Kit (Cat. No. 69504) following a user-developed protocol ( https://www.studocu.com/en-au/document/la-trobe-university/bio-business-management/dy04-purification-of-total-dna-from-nails-hair-or-feathers-using-the-dneasy-blood-tissue-kit/9173223). Bisulfite conversion was carried out using the EZ DNA Methylation-GOLD™ Kit (Cat. No. D5005). Whole-genome bisulfite sequencing (WGBS) libraries were generated using the xGen Methyl-seq Library Prep Kit (10009824, IDT). NEBNext Enzymatic Methyl-seq Kit (E7120) and or xGen Methyl-seq Library Prep Kit (10009824, IDT)