GSM6627989: Aorta, wild type, rep1; Mus musculus; RNA-Seq
Aorta were incubated with Trizol reagent and centrifuged at 13000×g for 5 min at 4℃. The supernatant was collected and mixed with chloroform. After centrifugation, RNA in the aqueous phase was precipitated by mixing with isopropanol at -20℃ for 1 h. Then, the RNA pellet was washed for two times and resuspended. The sample was centrifuged at 13000×g for 5 min at 4℃. The dried pellet was dissolved by DEPC-treated water. The quality and quantity of total RNA were assessed using Nano Drop  and Agilent 2100 Bioanalyze. The mRNA was purified and fragmented into small pieces. Then first-strand cDNA was generated by random hexamer-primed reverse transcription, followed by a second-strand cDNA. The cDNA fragments were amplified by PCR, and the PCR products were purified by Ampure XP Beads. The purified PCR produces were heated denatured and circularized by the splint oligo sequence to get the final library. The final library was amplified with phi29 to make DNA nanoball (DNB) which had more than 300 copies of one molecular. The DNBs ware loaded into the patterned nanoarray