GSM5706142: HEAP miRNA library, Halo-Ago2/+; Fabp-Cre/+, rep 1 (HF_miRNA1); Mus musculus; RIP-Seq
Distal colon was scraped using a razor blade to enrich for colonic epithelium. Tissue was minced on ice and UV-crosslinked (400mJ/cm2, 3X). Scraped tissues were flash frozen in liquid nitrogen and stored at -80ºC. Frozen samples were powderized in liquid nitrogen with a tissue homogenizer before library prep. Published protocl was followed (PMID: 32497496). In short, Halo-Ago2/mRNA/miRNA complex was pulled-down from tissue lysate. Ago2 was digested to release miRNA and mRNA. RNA was then used for library construction. RNA extracted from Input lysate without Halo-Ago2 pulldown was used for Input Control library construction. HEAP (CLIP-Seq)