title: GSM1181985: Caco-2 RNC-mRNA; Homo sapiens; RNA-Seq
library_construction_protocol: See the paper " Translating mRNAs strongly correlate to proteins in a multivariate manner and their translation ratios are phenotype specific" Wang et. Al. NAR 2013. Briefly, the polyA+ mRNA in the total mRNA or RNC-mRNA samples was isolated using the RNA Purification Beads (Illumina). The mRNA was fragmented by incubation in Elute-Prime-Fragment Mix at 94°C for 8 min to obtain 120–200 bp inserts See the paper " Translating mRNAs strongly correlate to proteins in a multivariate manner and their translation ratios are phenotype specific" Wang et. Al. NAR 2013. Briefly, the polyA+ mRNA in the total mRNA or RNC-mRNA samples was isolated using the RNA Purification Beads (Illumina). The mRNA was fragmented by incubation in Elute-Prime-Fragment Mix at 94°C for 8 min to obtain 120–200 bp inserts. First-strand cDNA was synthesized with SuperScript II Reverse Transcriptase (Invitrogen) using random primer, and Ampure XP beads (Beckman Coulter, Beijing, China) were used to isolate double-stranded cDNA synthesized by Second Strand Master Mix. The adapters were ligated to the A-Tailing fragment, and 12 cycles of PCR were performed to enrich those DNA fragments that have adapter molecules on both ends and to amplify the amount of DNA in the library. Purified libraries were quantified by Qubit® 2.0 Fluorometer (Invitrogen) and validated by Agilent 2100 bioanalyzer (Agilent, Beijing, China). Clusters were generated by cBot with the library diluted to 10 pM