Abstract

Selfie-qPCR : accurate expression measurement without reference gene

Petar Podlesniy, Postdoctoral Research Associate, Consejo Superior De Investigaciones Científicas

From the early years of the qPCR the estimation of the gene expression regulation is based on comparison with a reference, also known as housekeeping or normalization gene considered being a stable value. No such a gene applicable to wide range of species and experimental problems is found up to date creating the necessity of multiple reference genes or other normalization approaches which are just a partial solution. We would like to propose a very simple and effective absolute gene expression level estimation and to demonstrate its applicability to a range of experimental problems. Selfie-qPCR is an approach for accurate estimation of absolute number of transcripts per genome copy or cell which we propose as a methodology which will permit obtaining a comparable absolute/digital expression levels between laboratories, experiments and organisms. Petar Podlesniy is a postdoctoral fellow of CiberNed in the Institute of Biomedical Research in Barcelona. He is an early adopter of ddPCR technology in the field of circulating cell-free nucleic acid as biomarkers of neurodegenerative diseases. His current research interest is dedicated to the development of direct qPCR/ddPCR approach for the interrogation of biological systems.