Reliability of RT-qPCR from degraded RNA samples: An in vitro model

Abstract A rapid and reliable protocol was developed to produce chemical degradation of the RNA in vitro. Cell line MDA-MB-453 was used as source of RNA and hydrolysis in water at 70°C was performed. The outcome was monitored by RT-qPCR of two housekeeping targets and a significant correlation between the Ct and the extent of molecular damage of the template was found. A molecular degradation index of the RNA is proposed.