GSM8942706: Pituitary, CCK1/2-KO, male, rep1; Danio rerio; RNA-Seq
Pituitary  tissues were dissected, washed in ice-cold minimal essential medium, and dissociated using trypsin, EDTA, and DNase II. After cell counting, cells were washed twice in NeuroGro medium (T710KJ, BasalMedia, Shanghai, China) and resuspended in PBS at a concentration of 1 × 10⁶ cells per mL The 10X Genomics was used to generate single-cell Gel Beads-in-emulsion (GEMs). Immediately after GEM generation, the gel bead was dissolved, primers including an Illumina TruSeq Read 1 (read 1 sequencing primer), 16 nt 10x barcode, 12 nt unique molecular identifier (UMI), 30 nt poly(dT) sequence were released and mixed with cell lysate and a master mix containing reverse transcription (RT) reagents. Silane magnetic beads were used to purify first-strand cDNA from the post-GEM-RT reaction mixture, which contained residual biochemical reagents and primers. Barcoded full length cDNA was amplified by PCR to generate sufficient mass for library construction.