GSM5709569: GATA2_KO1; Mus musculus; RNA-Seq
RNA extraction was performed according to manufacturers' instructions using the NucleoSpin RNA XS kit (cat # 740902, Macherey-Nagel). Briefly, FACS isolated gonadotrope cells were lysed in 100ul of lysis buffer (RA1 reagent + TCEP), DNA was digested with rDNAse for 15min at RT, and the final RNA eluted in 10ul of RNAse-free water. RNA concentration was measured by Nanodrop-One (Thermo Scientific), and the RNA quality was assessed on a Bioanalyzer (cat# 5067-1513, Eukaryote Total RNA Pico Bioanalyzer kit, Agilent). RNAseq libraries were prepared with the extracted RNA using the Universal plus mRNA-seq library preparation kit (NuGEN). Library quality control and quantification were assessed by fluorometry (cat # Q32854, Qubit dsDNA High sensitivity Assay Kit), and on a Bioanalyzer (cat # 5067-4626, High-Sensitivity DNA Bioanalyzer kit, Agilent). The 4 libraries were pooled together at equal concentration, the pooled sample was first assessed by mi-seq (Illumina), and then sequenced at the New York Genome Center on Novaseq 6000 (Illumina) with a depth of ~30 million paired-end reads per sample. 