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

Authors :: Paola Pitacco
Vanessa Nicolin
Giorgio Marrubini
Paolo Vatta
Carlo Previderè
S. Sorçaburo Cigliero
Serena Bonin
R. Palladino
Paolo Fattorini
Palladino, R.
Cigliero, S. Sorçaburo
Bonin, Serena
Pitacco, P.
Nicolin, Vanessa
Previderè, C.
Vatta, P.
Marrubini, G.
Fattorini, Paolo
Source :: Forensic Science International: Genetics Supplement Series. 5:e60-e62
Publication Year :: 2015
Publisher :: Elsevier BV, 2015.
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.