GSM9765361: Lgr5-GFP+ ISCs, KANSL1-KO, day4, rep2; Mus musculus; RNA-Seq
Total RNA was extracted using the MJzol Animal RNA Isolation Kit (Majorivd) following the manufacturer's standard protocol, and purified using the RNAClean XP Kit (Beckman Coulter) and the RNase-Free DNase Set (QIAGEN). RNA integrity was assessed using the Agilent 2100 Bioanalyzer/Agilent 4200 TapeStation (Agilent Technologies), while total RNA yield and purity were determined using the Qubit 2.0 Fluorometer (Thermo Fisher Scientific) and the NanoDrop ND-2000 Spectrophotometer (Thermo Fisher Scientific). The purified total RNA was subjected to mRNA isolation, fragmentation, first-strand cDNA synthesis, second-strand cDNA synthesis, end repair, 3′ end A-tailing, adapter ligation, and enrichment to complete mRNA sequencing library construction. The library concentration was measured using the Qubit 2.0 Fluorometer (Thermo Fisher Scientific), and the library fragment size distribution was assessed using the Agilent 4200 TapeStation (Agilent Technologies).