GSM9131347: Kcnj2 KO/KO 2 13.5 anterior palate; Mus musculus; RNA-Seq
In ice-cold PBS, heads were removed from embryos. The top of the head was cut flat, and the head was laid ventral. The secondary palate was exposed by removing the mandible and tongue. The anterior 1/3 of the palate was dissected away from the remainder of the head using forceps. Paired anterior palatal shelves were washed twice in PBS and transferred to 750 uL 0.25% Trypsin-EDTA for 10 minutes at 37C with frequent trituration. Trypsinization was stopped with addition of 750uL DMEM + 10% FBS . Cells were pipetted up and down 20 times to assist in cell dispersion, pelleted at 300 g for 3 min, and resuspended in PBS with 0.1% BSA before being filtered through a Flowmi 40uM cell strainer. Single cell suspensions were confirmed by microscopy and cells were counted on a hemocytometer with trypan blue staining. Chromium single cell 3' NextGen V3.1 chemistry (10X Genomics)