GSM6615245: siGAPDH [si-14-p_S66]; Homo sapiens; RNA-Seq
Qiagen RNAeasy kits to extract total RNA from hPAEC cell lysates from confluent 6-well plates. Each RNA sample will be quantified using Qubit and run on a Tapestation to assess sample integrity. mRNA will be isolated from total RNA using oligo dT beads. After fragmentation the sample will be converted to double stranded DNA and ligated to Illumina adapters. The second strand cDNA is then degraded by the UNG enzyme providing directional information to the library. Indexing of each sample will be performed by PCR. For each pool of libraries, Tapestation will be used to assess fragment distribution and presence of artefacts. Each submitted pool will also be quantified by Qubit and/or qPCR prior to sequecing. Sequencing will be performed using a HiSeq4000. pooled and sequenced across multiple lanes