GSM4566290: D3_N_T_2; Homo sapiens; RNA-Seq
Total RNA was isolated using using the Norgen Biotek kit-QC before library construction was carried out via: (1) Nanodrop: for RNA purity (OD260/OD280).  (2) Agarose gel electrophoresis: for RNA integrity and potential contamination.  (3) Agilent 2100 Library preparation was done using the  NEB Next® Ultra™ RNA Library Prep Kit. After the QC procedures, mRNA from eukaryotic organisms is enriched using oligo(dT) beads. First, the mRNA is fragmented randomly by adding fragmentation buffer, then the cDNA is synthesized by using mRNA template and random hexamers primer, after which a custom second-strand synthesis buffer (Illumina) , dNTPs, RNase H and DNA polymerase I are added to initiate the second-strand synthesis. Second, after a series of terminal repair, A ligation and sequencing adaptor ligation, the double-stranded cDNA library is completed through size selection and PCR enrichment. Double stranded cDNA synthesis, end repair, poly A/Adaptor addition, fragment selection, PCR
GEO Accession
GSM4566290