title: GSM9656263: C1285_insC_3: FLCN c.1285_1286insC (p.H429Pfs*27) mutant plasmid, biological replicate 3; Homo sapiens; RNA-Seq
library_name: GSM9656263
library_construction_protocol: Total RNA was isolated from cultured Nthy-ori 3-1 cells (Mock control, FLCN wild-type transfected, FLCN-siRNA knocked down, and FLCN mutant (c.992C>T, c.1179delC, c.1285_1286insC) transfected groups) using a standard TRIzol/chloroform extraction method (or commercial RNA extraction kit). RNA purity and integrity were assessed by Nanodrop and Agilent 2100 Bioanalyzer (RNA Integrity Number, RIN ≥ 8.0 was required for library construction). cDNA libraries were constructed from qualified total RNA using the Illumina TruSeq Stranded Total RNA Library Prep Kit (Human/Mouse/Rat) (or Personalbio customized library construction kit) following the manufacturer's instructions. Ribosomal RNA (rRNA) was removed prior to cDNA synthesis to enrich for coding and non-coding RNA. The RNA was fragmented, reverse-transcribed into first-strand cDNA, and second-strand cDNA was synthesized with dUTP to achieve strand specificity. cDNA fragments were end-repaired, A-tailed, and ligated with Illumina adaptors, followed by PCR amplification to generate the final cDNA libraries.
identifiers.namespace: GEO
identifiers.id: GSM9656263_r1