GSM1697526: E5_C4_24h; Homo sapiens; RNA-Seq
Trizol extraction of total RNA was performed according to the manufacturer's instructions Total RNA was subjected to two rounds of poly A+ selection and converted to cDNA. We used TruSeq library generation kits as per the manufacturer’s instructions (Illumina, San Diego, CA, USA). Library construction consisted of random fragmentation of the polyA mRNA, followed by cDNA production using random primers. The ends of the cDNA were repaired, A-tailed and adaptors ligated for indexing (up to twelve different barcodes per lane) during the sequencing runs. The cDNA libraries were quantitated using qPCR in a Roche LightCycler 480 with the Kapa Biosystems kit for library quantitation (Kapa Biosystems, Woburn, MA) prior to cluster generation. Clusters were generated to yield approximately 725K-825K clusters/mm2. Cluster density and quality was determined during the run after the first base addition parameters were assessed. We ran paired end 2X50bp sequencing runs to align the cDNA sequences to the reference genome
GEO
GSM1697526
GEO Accession
GSM1697526