title: GSM3146360: H-fibroblast; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNAs were isolated using Trizol Reagent (Gibco). 1-μg RNA samples were reverse transcribed using a Transcriptor First Strand cDNA Synthesis Kit (Roche, PA, USA). A library was independently prepared with 1 µg of total RNA for each sample using an Illumina TruSeq Stranded mRNA Sample Prep Kit (#RS-122-2101, Illumina, Inc., San Diego, CA, USA). The initial step of library preparation involved purifying poly‐A containing mRNA molecules using poly‐T-attached magnetic beads. After this step, the purified mRNA was fragmented into small pieces using divalent cations under elevated temperature. The cleaved mRNA fragments were copied to the top strand of cDNA using SuperScript II reverse transcriptase (Invitrogen, #18064014) and random primers, and the reverse complement strand of cDNA was synthesized using DNA polymerase I, RNase H, and dUTP. These cDNA fragments underwent the ends repair process including the addition of a single 'A' base and adapter ligation. After these steps, the final cDNA libraries were created, purified, and enriched with PCR
attributes.tag: GEO Accession
attributes.value: GSM3146360