GSM8385431: Python bivittatus left ventricle, Fasted 2; Python bivittatus; RNA-Seq
Frozen ventricular tissue was homogenized in Trizol and then incubated at room temperature for five minutes. Chloroform was added at 1:5 (v/v), the tubes shaken vigorously, and incubated at room temperature for 15 minutes. Samples were centrifuged at 12,000 RCF for 15 minutes and the upper aqueous layer collected and added to an equal volume of isopropanol. Then samples were incubated at -20 °C for 15 minutes and then centrifuged at 12,000 RCF for 10 minutes to pellet the RNA. The RNA pellet was washed twice with ice-cold 75% ethanol and solubilized in RNase-free water. RNA was sent to Novogene Corporation (Sacramento, CA) for mRNA enrichment and short-read RNA sequencing at a depth of 50 million paired-end reads per sample.