GSM4477472: H1_hESC_KLF4KO_derived_hPGCLCs_p68; Homo sapiens; RNA-Seq
Total RNA was extracted from H1_hESC_WT_derived_hPGCLCs and H1_hESC_KLF4KO_derived_hPGCLCs at passage days 65 and 68, using RNeasy micro kit (Qiagen 74004). Total RNA was reverse transcribed and cDNA was amplified using Nugen RNA-Seq System V2 (Nugen, 7102-32). DNA was extracted using MinElute PCR purification kit (Qiagen) and quantified using the Qubit dsDNA High-Sensitivity Kit (Life Technologies).  Amplified cDNA was fragmented using Covaris S220 Focused-ultrasonicator. RNA-sequencing libraries were generated using Nugen Rapid Library Systems (Nugen 0320-32). Libraries were subjected to single-end 125 bp sequening on HiSeq4000 with 6 indexed libraries per lane.