GSM5706165: HEAP IC library, Halo-Ago2/+; Villin-CreER/+; Apc(fl/fl), rep 2 (HVA_IC2); Mus musculus; RIP-Seq
Animals were sacrified when they became moribund.Tumor lawns along colon were resected or scraped using a razor blade. 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)