GSM8137001: High BMI sample 1; Homo sapiens; RNA-Seq
The fresh biopsy intestinal fragments from 2-3 volunteers were mixed into one tube, incubated with EDTA buffer, suspended and washed in cold PBS to obtain the crypt fractions as described above. Followed by shaking-incubated with digestive solution containing 1TrypLE Express (Gbico, 12604021) and 0.2 mg/ml DNase I (Thermo, EN0523) at 37℃ for 3-6 minutes, the single cells were centrifuged at 4℃ with 350g for 5 minutes. Then the cell pellets were resuspended and filtered through a 40 m cell strainer (BD). After centrifugating and discarding the supernatant, the cell sediments were stained with Zombie Dyes (Biolegend, 423105; 1:100 dilution) in the dark at the room temperature for 10 minutes. Zombie-negative cells were then sorted and collected by FACS analysis (BD, melody) Single cells were captured in the 10X Genomics Chromium Single Cell 3′ Solution, and RNA-seq libraries were prepared according to the manufacturer's protocol (10X Genomics). Then the libraries were sequenced on an Illumina Hiseq X Ten PE150 platform.