GSM5620448: HUVECs treated with HG+IS 30IS-1_L3_A021; Homo sapiens; RNA-Seq
Total RNA was extracted from cultured cells using NucleoZOL reagent according to the manufacturer's protocol. One μg of total RNA was reverse-transcribed by a High-Capacity cDNA Reverse Transcription Kit RNA concentration and quality were determined using a Qubit Fluorometer (Thermo Fisher Scientific, USA) and an Agilent 2100 Bioanalyzer which calculates an (RIN).The ribosomal RNAs (rRNA) were eradicated from total RNA using a Ribo-zero rRNA removal kit (Cat. no. 20278677, Illumina, USA). After purification, the RNAs were fragmented into small pieces by using divalent cations with elevated temperature. The cleaved RNA fragments were copied into first strand cDNA using reverse transcriptase and random primers. Followed by second strand cDNA synthesis using DNA Polymerase I and RNase H. cDNA fragments went through an end repair process with addition of a single “A” base and ligation of the adapters.The products were then purified and enriched with PCR to create the final cDNA library.Paired-end reads from each cDNA library were obtained using a NextSeq500 system (Illumina Inc., USA).
GEO Accession
GSM5620448