title: GSM8241979: E13.5_PGC_Control_Female_2; Mus musculus; RNA-Seq
library_construction_protocol: PGCs from dissected embryonic gonads (E12.5, E13.5, E14.5) and dissected embryo trunks (E10.5, E11.5) were dissociated in 0.05% Trypsin in 5% CO2/37C for 5 minutes. Digestions was quenched using 10% FBS in DMEM High Glucose media and cells were pelleted at 1200 rpm x 5 minutes. Resulting pellet was resuspended in 10% FBS, High Glucose DMEM + DNAse and treated for 5 minutes at 5%CO2/37C and pelleted by centrifugation at 1,200 RPM for 5 minutes. PGCs were then suspended in DPBS + 1% BSA and GFP+, 7AAD- cells were collected in Qiagen Buffer RLT. RNA was isolated with Qiagen RNEasy kit according to manufacturers instructions. cDNA conversion was performed using Tecan Genomics Ovation RNA-seq System V2. Resulting cDNA was sonicated using a Covaris S2 sonicator and the resulting fragmented cDNA isolated using Quiagen MinElute columns according to the manufacturer's instructions. Isolated cDNA was repaired and barcoded using Tecan Genomics Encore Rapid Library barcoding according to the manufacturers instructions. Samples were quantified using KAPA quanitifcation according to the manufacturer's instructions. Libraries were sequenced using a NovaSeq 6000.