GSM2425076: SSEA4+_SSCs_patient2_rep1; Homo sapiens; RNA-Seq
RNA was extracted from pooled SSEA4+ or Kit+ germline stem cells using AllPrep RNA/DNA/Protein Mini Kit (QIAGEN cat # 80004). Total RNA was then subjected to RiboZero Gold (Illumina cat # MRZG126) to substantially deplete cytoplasmic and mitochondrial rRNA. Total RNA samples (100-500 ng) were hybridized with Ribo-Zero Gold to substantially deplete cytoplasmic and mitochondrial rRNA from the samples.  Stranded RNA sequencing libraries were prepared as described using the Illumina TruSeq Stranded Total RNA Kit with Ribo-Zero Gold (RS-122-2301 and RS-122-2302).  Purified libraries were qualified on an Agilent Technologies 2200 TapeStation using a D1000 ScreenTape assay (cat# 5067-5582 and 5067-5583).  The molarity of adapter-modified molecules was defined by quantitative PCR using the Kapa Biosystems Kapa Library Quant Kit (cat#KK4824).  Individual libraries were normalized to 10 nM and equal volumes were pooled in preparation for Illumina sequence analysis. Libraries were then sequenced on a 50-cycle single end run on an Illumina HiSeq 2500 instrument.
GEO Accession
GSM2425076