title: GSM4066257: Profiling of RNA from rRNA-depleted RNA library from primary tumors 3; Homo sapiens; RNA-Seq
library_construction_protocol: Tumor samples (PT and LM) were cutted into smaller pieces before putting them into the lysing matrix tubes, type "D" (MP Biomedicals). Tubes with 1 ml TRIzol (Invitrogen) were cooled on ice before samples were added. Tumors were homogenized using Fastprep-24-System with 1x 45 sec. @ full speed in TRIzol. After homogenisation, total RNA from tumor samples was isolated according to TRIzol's protocol. Total RNA from HT29 cells was isolated using TRIzol according to TRIzol's protocol without homogenisation steps. Workflow consists of double-stranded cDNA generation using a customizable mixture of random and poly(T) priming, fragmentation of double-stranded cDNA, end repair to generate blunt ends, adaptor ligation, strand selection via nucleotide analog-targeted degradation, InDA-C-mediated adaptor cleavage and PCR amplification to produce the final library. The detailed protocol can be found at the user guide Ovation Human FFPE RNA-Seq System.
attributes.tag: GEO Accession
attributes.value: GSM4066257