GSM7882412: Sample4_asthmatic model1; Rattus norvegicus; RNA-Seq
Total RNA was extracted and purified by the mirVana™ miRNA Isolation Kit (catalog number: AM1561; Ambion, Austin, TX, USA) according to manufacturer instructions. RNA quality was evaluated by a spectrophotometer (Nanodrop™; Thermo Fisher Scientific, Waltham, MA, USA) and a bioanalyzer (2100 series; Agilent Technologies, Santa Clara, CA, USA). The mRNA with polyA structure in the total RNA was enriched by Oligo(dT) magnetic beads, and ion interruption was used to break the RNA to a fragment of about 300bp in length. The fragment with a length of 300bp was chosen because the length of the junction is fixed, and if the length of the interrupted fragment is short, it will lead to a high proportion of the junction sequence, which will reduce the proportion of valid data; if the length of the interrupted fragment is long, it will be unfavorable to the generation of clusters in the process of on-line sequencing. The first strand of cDNA was synthesized using RNA as template with 6-base random primer and reverse transcriptase, and the first strand cDNA was used as template for the second strand cDNA synthesis.