GSM7900380: Longissimus dorsi muscle of Suhuai pig; Sus scrofa; RNA-Seq
The 1-day-old Suhuai boar was anaesthetized before the trial started. Then, the skin of the back was disinfected, and the longest dorsal muscle tissue was taken for cell suspension preparation using a sterile scalpel (1 cm3).The collected tissues were washed 3 times with PBS, the appropriate amount of PBS was added, and the tissues were cut with ophthalmic scissors (<1 mm). Add 5 ml of tissue lysate and incubate at 37°C. The tissue digest contained 4 mg/ml papain (Sigma‒Aldrich, cat# 9001-73-4), 20 mg/ml collagenase II (Sigma‒Aldrich, cat# 9001-12-1), and 20 mg/ml collagenase IV (Sigma‒Aldrich, sku# C4-22-1G). The digestion reactions were shaken vigorously for 30 s and further incubated at 37°C for 30 to 40 min in an incubator with general shaking every 6 min to release cells. The released cells were passed through a 70-μm cell strainer (BD, 352350) and collected in 15-mL tubes. Sample viability was assessed via Trypan Blue (Thermo Fisher) and an automatic cell counter (Countstar). GEMs are generated by combining barcoded Single Cell 3' v3.1 Gel Beads, a Master Mix containing cells, and Partitioning Oil onto Chromium Next GEM Chip G. Barcoded, full-length cDNA is amplified via PCR to generate sufficient mass for library construction.P5, P7, i7 and i5 sample indexes, and TruSeq Read 2 (read 2 primer sequence) are added via End Repair, A-tailing, Adaptor Ligation, and PCR. The final libraries contain the P5 and P7 primers used in Illumina amplification.