GSM2462054: FibroblastT45Cntrl 3; Homo sapiens; RNA-Seq
At 45, 60, 75 and 90 minutes post-infection, RNAprotect Cell Reagent (Qiagen) was added. Cells were harvested by scraping, pooled and pellets were resuspended in RLT-plus buffer (Qiagen) and stored at -80C. Cells were lysed with 27G1/2 needles before homogenization with QiAshredder columns (Qiagen). Total RNA was then purified using the RNAeasy Plus Kit (Qiagen). RNA concentration was determined with a Qubit fluorimeter prior to library prep.  4μg of total DNase treated RNA were prepared with the TruSeq Stranded Total RNA LT Sample Prep Kit from Illumina. In brief, polyadenylated RNA was purified and fragmented before strand specific cDNA synthesis.  cDNA was A-tailed and indexed adapters were ligated. After adapter ligation, samples were PCR amplified and purified with AmpureXP beads, then validated again on the Agilent 2100 Bioanalyzer.
GEO Accession
GSM2462054