GSM5335488: Single Cell RNA-Seq of the Male Embryonic Day 14.5 Genital Tubercle_1_1; Mus musculus; RNA-Seq
The genital tubercles were then dissected from the body and placed into 1.3mg/ml collagenase XI (Sigma-Aldrich, C7657-25MG) on ice. Collagenase XI was made by first dissolving 13mg into 1ml TESCA buffer at 37°C for 5 minutes and then further diluting to 1.3mg/ml in 10% inactivated fetal bovine serum/ Dulbecco's phosphate-buffered saline. The genital tubercles were chopped into small pieces using forceps while in 500μl collagenase XI on ice. The entire 500μl of collagenase XI and tissue was collected and placed into a 1.5ml microcentrifuge tube for remainder of digestion.  Cells were agitated at 37°C for 30 mins and tritrated with a glass pipettes every 5 mins. After digestion, cells were centrifuged at 500rpm for 5 minutes at 4°C, then were was resuspended in 100ul of magnesium and calcium free 1xPBS with 0.04% bovine serum albumin, filtered through a 40μm strainer (Flowmi Bel-Art H13680-0040), and placed on ice.