GSM3755319: disp_rep2; Homo sapiens; RNA-Seq
High quality total RNA was extracted from samples using Nucleospin RNA kit of Macherey-Nagel and eluted in RNAse free water. RNA-Seq library preparations were done using established Illumina methods for mRNA-Seq (Part #RS-  122-2001). Briefly, poly A+ RNA was purified from 100 ng of total RNA with oligo-dT beads. Purified  mRNA was fragmented with divalent cations at elevated temperature, to ~200bp. First strand cDNA  synthesis was performed with random hexamer priming and SuperScript II Reverse transcriptase  (Invitrogen, part # 18064-014). Second strand cDNA synthesis was performed using RNAseH and DNA  PolI. Following dscDNA synthesis, the double stranded products are end repaired, followed by addition of  a single 'A' base and then ligation of the Illumina TruSeq adaptors. The resulting product was amplified  with 15 cycles of PCR. Each library was made with a unique Index sequence and libraries are pooled at the  desired plexity. The pools were clustered at 6.5pM on a single end read flow cell and sequenced for 50  cycles on an Illumina HiSeq 2500. Primary processing of sequencing images was done using Illumina's  Real Time Analysis software (RTA) as suggested by the Illumina. CASAVA 1.8.2 software was then used  to demultiplex samples and generate raw reads and respective quality scores. Illumina's CASAVA 1.8.2  software was used to perform image capture, base calling and demultiplexing.
GEO Accession
GSM3755319