GSM9534403: PGC_D125, rep1; Gallus gallus; RNA-Seq
Embryonic gonads at E7, E8, E9, E10, and E11 were digested with 100 µL trypsin (0.05%), followed by addition of 900 µL of Dulbecco's Modified Eagle's Medium containing 10% fetal bovine serum. Then the samples were centrifuged at 200 × g for 5 min and resuspended in 1mL PBS. After using 40 μm sterile strainers to filter, the samples were centrifuged at 350× g for 5 min again, pelleted down and resuspended in PBS. Single-cell suspensions were carried out with 10x Genomics Chromium Single Cell Kit for Single-cell RNA-seq. single-cell suspensions with PBS were loaded onto microwell chip using the Singleron Matrix® Single Cell Processing System. Barcoding Beads were subsequently collected from the microwell chip, followed by reverse transcription of the mRNA captured by the Barcoding Beads to obtain cDNA, and PCR amplification. The amplified cDNA was then fragmented and ligated with sequencing adapters. The scRNA-seq libraries were constructed according to the protocol of the GEXSCOPE® Single Cell RNA Library Kits (Singleron).