GSM5925135: adult_rep_2; Danio rerio; RNA-Seq
Adult and aged zebrafish were anesthetized on ice. The whole brain was dissected in ice-cold PBS and collected into buffer (fresh PBS with 5% fetal bovine serum (FBS) and 0.1% DNaseI). The collected brains was cut into pieces, dissociated with digested buffer (0.25% trypsin in buffer) for 15 min and passed through a 26G needle 5 times to further smash the tissue. The processed cell samples were filtered with 40 uM cell strainer and collected with 800g at 4℃ for 5min, then resuspended to single-cell suspension with ice-cold 5% FBS. Next, cell sorting was performed by flow cytometry (Moflo XDP, Beckman) to gain GFP+ immunocytes.Then the collected single-cell products were performed 10X single cell RNA sequence RNA libraries were prepared with Chromium Single cell 3` Reagent Kits (v3.1 chemistry, PN- 1000121) according to the manufacture`s protocol