GSM5706158: HEAP mRNA library, Halo-Ago2/+; Fabp-Cre/+; K-Ras-G12D/+, rep 3 (HFK_mRNA3); 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)