alias: GSM4103045
title: GSM4103045: Cont-iEPCs; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNAs from Cont-iEPCs and YAC-iEPCs were purified using an Agencourt RNAdvance Tissue Total RNA Purification Kit (Beckman Coulter, Brea, CA, USA). Briefly, mRNA from Eukaryote organisms is purified from total RNA using poly-T oligo-attached magnetic beads (For prokaryotes, mRNA was purified through the removal of rRNA by using kit). The mRNA is first fragmented randomly by addition of fragmentation buffer. Then first strand cDNA is synthesized using random hexamer primer and M-MuLV Reverse Transcriptase (RNase H-). Second strand cDNA synthesis is subsequently performed using DNA Polymerase I and RNase H. Double-stranded cDNA is purified using AMPure XP beads. Remaining overhangs of the purified double-stranded cDNA are converted into blunt ends via exonuclease/polymerase activities. After adenylation of 3' ends of DNA fragments, NEBNext Adaptor with hairpin loop structure is ligated to prepare for hybridization. In order to select cDNA fragments of preferentially 150~200 bp in length, the library fragments are purified with AMPure XP system (Beckman Coulter, Beverly, USA). Finally, the final library is gotten by PCR amplification and purification of PCR products by AMPure XP beads. The library were preparated using NEBNext Ultra RNA Library Prep Kit for Illumina.
identifiers.namespace: GEO
identifiers.id: GSM4103045
attributes.tag: GEO Accession
attributes.value: GSM4103045