title: GSM8768873: Localized PCa patient 187; Homo sapiens; RNA-Seq
library_construction_protocol: RNA from 50ng of FFPERNAsamples.RNA from these samples were not fragmented previous to 1st strand cDNAsynthesis. The 1ststrand cDNA synthesis was performed using SMARTScribe reverse Transcriptase, for 90min. at 42°C, 10min. at 70°C and pause at 4°C. Afterwards Illumina Adapters and Indexes are added, performing a preamplification PCR (60sec at 94°, 5 cycles of 15sec at 98°C, 15sec a t55°C 30sec at 68°C; and pause at 4°C). Then Ribosomal cDNA is depleted with ZapR v2 and R-Probes v2. Finally, enrichment of libraries was achieved by PCR (60sec at 94°C; 13-19 cycles of 15sec at 98°C, 15sec at 55°C, 30sec at 68°C; and pause at 4°C). Sequencing libraries were prepared using “SMARTer Stranded TotalRNA-seq Kitv2–Pico Input Mammalian” kit (Takara Bio USA, Cat.# 634411), following “SMARTer Stranded Total RNA-seq Kitv2–Pico Input Mammalian User Manual (Rev.050619)”.