GSM3579942: Single cell RNAseq from adult male pituitary; Mus musculus; RNA-Seq
Mouse pituitary was dissected in 300µl of dissection buffer (DMEM, 10% FBS, HEPES 10mM and DNase 100 U/ml), dissociation was performed at 37°C using 5mg/ml Trypsin for 10 minutes. We then added 2mM EDTA and incubated 5 minutes more. Single cells preparation were obtained by filtration through 40µm filter. Dissociated pituitary cells were diluted at 500 cells per µl and processed using Chromium Single Cell 3′ v2 Reagent (10x Genomics, Pleasanton, CA) following the manufacturer recommendation. Briefly, cells were passed on the channel and 9269 cells were recovered. Pituitary cells were partitioned into gel beads in emulsion for cell lysis and barcoded with oligo-dT priming and reverse transcribed. cDNA library was amplified fragmented and size selected. Samples were controlled at multiple steps during the procedure by running on BioAnalyzer. Libraries were sequenced on Hiseq 4000 with 100bp paired-end reads.
GEO Accession
GSM3579942