GSM6304079: High_1_S13; Mus musculus; RNA-Seq
Collagenase (2mg/mL) and Dispase (5mg/mL) in PBS was injected beneath and beside the CVP, the epithelium peeled and then dissociated for 45 min in Collagenase (2mg/mL), Dispase (5mg/mL), and Elastase (2mg/mL) at 37°C. Cells were centrifuged (2000 RPM 4°C), the pellet resuspended in FACS buffer (1mM EDTA, 25mM HEPES pH 7.0, 1% FBS, 1x Ca2+/Mg2+-free dPBS), passed through a 30µm cell strainer and subjected to FACS for GFP signal. GFPneg cells were discarded PolyA selection was performed and sequencing libraries were synthesized using NuGen Universal Plus mRNA-Seq with NuQuant kit at the University of Colorado Genomics and Microarray Core. Libraries were sequenced on an Illumina NovaSeq6000.