title: GSM3461364: D16-1569_Yng RF pan (TACCGC); Mus musculus; RNA-Seq
library_construction_protocol: As previously reported and briefly summarized here, small intestines were removed, washed with cold PBS, opened longitudinally and then incubated on ice with PBS−/− EDTA (10 mM) for 30-45 min. Tissues were then moved to PBS. Crypts were then mechanically separated from the connective tis-sue by shaking, and then filtered through a 70-μm mesh into a 50-ml conical tube to remove villus material and tissue fragments. Crypts were removed from this step and dissociated into single cells and sorted by flowcytometry. PolyA+  RNA extraction (Spri beads), followed by RNA fragmentation, reverse transcription, end-repair and adjunction of Illumina adapters by PCR (TruSeq protocol, Spri size selection 200-400 bps).