GSM6212890: embryonic wt mouse palate e15.5 scRNA-seq; Mus musculus; RNA-Seq
GSM6212890
Embryonic palatal shelves were collected from the stages of E13.5, E14.5, and E15.5. Tissues from 3 embryos were pooled in 500 µL PBS in 0.04% BSA in a 1.5 mL tubes on ice. Centrifuged the tubes with tissue using mini centrifuge for few seconds. Removed the supernatant and cut the tissues into small fragments using a small surgical scissor. Added 1mL of PBS in 0.04% BSA, spin the tissue down using mini centrifuge and removed supernatant. Enzymatic digestion of the tissues was done using TrypLE express enzyme at 37°C with shaking at 600 rpm for 20 minutes. Occasional pipetting was done with a wide bore pipette tip. Added 4mL DMEM medium to stop enzymatic digestion and mixed well by gentle pipetting. Filtered the cells using a 70 µM cell strainer and centrifuged at 1500 rpm for 5 minutes. Removed supernatant and added 5mL of PBS in 0.04% BSA into the cell pellet and centrifuged at 1500 rpm for 5 minutes. Removed the supernatant and single cell suspension was fixed in methanol according to 10X Genomics protocol and stored in -80°C. Libraries were prepared according to Illumina Single cell RNA seq preparation procotol