GSM1695260: hydrostatic pressure plus perfusion replicate1; Homo sapiens; RNA-Seq
After 4 days of culture, total RNA was isolated using QiazolTM Lysis Reagent (Qiagen, Hilden, Germany) according to manufacturer’s introduction An amount of 100 ng total RNA was first treated with double-strand specific DNase (Fermentas Inc., Hanover. MD , United States) to remove any traces of genomic DNA. After heat-inactivation of the DNase, cDNA was synthesized and converted to Illumina-compatible sequencing libraries with the Encore complete RNAseq kit from NuGen (NuGen, San Carlos, USA). Briefly, first strand cDNA was generated by selective priming, second strand cDNA was synthesized using dUTP and the generated double stranded cDNA was fragmented using a Covaris M220 sonicator (50W peak incident power, 20% duty factor, 200 cycles per burst, 160 sec treatment time). Then cDNA was end-repaired, ligated to Illumina-Adapters and the second-strand was selectively removed. After 18 cycles of PCR the final library was quantified
GEO Accession
GSM1695260