GSM4061872: KIS1 EBF1 - DOX 2; Homo sapiens; RNA-Seq
RNA was extracted with trizol and subsequently the RNAeasy minikit RNA was quantified and integrity was analyzed on Agilent Tape Station. Samples with RNA integration number of greater than 8 were selected for library preparation. Poly-A enrichment of mRNA was performed from 15ug of total RNA (Dyna beads mRNA direct Micro Kit). All samples were spiked in with ERCC spike in mix (Life Technologies) before poly-A enrichment for performance assessment of libraries. 100ng of poly-A enriched RNA was fragmented using RNase III and purified using magnetic bead clean up module (RNA Seq V2 kit, Life Technologies). Size distribution of the fragmented RNA was assessed on Agilent Tape Station using RNA HS screen tape. 50ng of fragmented poly A enriched RNA was used to prepare whole transcriptome library using RNA seq V2 kit( Life Technologies ) following manufacturer recommended protocols . Yield and size distribution of the  library was assessed on Agilent Tape Station using D1000 screen tape. Barcoded libraries were equally pooled and amplified onto Ion Sphere™ Particles (ISPs) supplied by Ion Pi HiQ OT2 kit (Life Technologies). ISPs loaded with libraries were sequenced on Ion Torrent Proton sequencer using Ion PI chip V3. Two samples were pooled together and sequenced in one chip (Ion PI HiQ sequencing kit).
GEO
GSM4061872
GEO Accession
GSM4061872