GSM3384195: CMT-077-tumor; Canis lupus familiaris; RNA-Seq
Fresh tissue samples were transferred in to RNAlater (Thermo Fisher Scientific), refrigerated overnight at 4°C and then stored at -80°C. Total RNA was extracted from tissues using RNeasy mini kit (Qiagen). RNA quality was assessed by analysis of rRNA band integrity on an Agilent RNA 6000 Nano kit. Ahead of cDNA library construction, the 1ug of total RNA and magnetic beads with Oligo (dT) were used to enrich poly (A) mRNA from it. Then, the purified mRNAs were disrupted into short fragments, and the double-stranded cDNAs were immediately synthesized. The cDNAs was subjected to end-repair, poly (A) addition, and connected with sequencing adapters using the TruSeq RNA sample prep Kit (Illumina, CA). The suitable fragments automatically purified by BluePippin 2% agarose gel cassette (Sage Science, MA) were selected as templates for PCR amplification. The final library sizes and qualities were evaluated electrophoretically with an Agilent High Sensitivity DNA kit (Agilent Technologies, CA) and the fragment was found to be between 350–450 bp. Libraries were sequenced on the HiSeq 2500 following the manufacturer's protocols.
GEO
GSM3384195
GEO Accession
GSM3384195