title: GSM8760147: Apc flox;Tcf7 del biol rep 1; Mus musculus; RNA-Seq
library_name: GSM8760147
library_construction_protocol: Small intestines of Apcfl/fl mice were cut open, washed with PBS, and incubated in 10 mM EDTA in a shaker at 37°C for 10 min. Tissues were washed twice with PBS, and the cells were scraped off with a coverslip. The cells were then centrifuged at 1500 RPM for 5 min, followed by incubation with a digestion mix composed of HBSS with Mg2+ and Ca2+ (Gibco, #14025-050) and supplemented with 1 mg/ml of collagenase type I (Gibco,#17100-017), collagenase type IV (1 mg/ml) (Gibco #17104-019), 4 mg/ml of Dispase II (Sigma Aldrich, #04942078001), and 1 µl/ml of DNase1 (Thermo Fisher Scientific, #EN0521) for 30-40 min at 37°C. After this, DMEM was added, and the cell suspension was filtered through 70 µm and twice through a 40 µm filter and centrifuged for 10 min at 150 RCF. TdTomato+ cells from the mice were sorted using BD Influx (BD Biosciences). Sorted cells were resuspended in HBSS containing 0.04% BSA and subjected to Chromium X. Library was performed according to the manufacturer`s instructions (single cell 3' protocol, 10x Genomics)