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JMD 2006, Vol. 8, No. 5
Copyright © 2006 American Society for Investigative Pathology & Association for Molecular Pathology

Technical Advance

Betaine, Dimethyl Sulfoxide, and 7-Deaza-dGTP, a Powerful Mixture for Amplification of GC-Rich DNA Sequences

Marco Musso^*, Renata Bocciardi^*, Sara Parodi^*, Roberto Ravazzolo^* and Isabella Ceccherini^*

From the Laboratory of Molecular Genetics, * G. Gaslini Institute, Genoa; and the Departments of Pediatrics & Centre of Excellence for Biomedical Research, University of Genoa, Genoa, Italy

Abstract

Currently, polymerase chain reaction is the most used technique in many laboratories for either diagnostic or molecular biology purposes. Despite the large number of DNA sequences that can be easily analyzed, some GC-rich sequences are refractory to amplification due to the formation of secondary intramolecular structures. To overcome this problem, several molecules have been described to improve polymerization. Here we show that a combination of three additives—betaine, dimethyl sulfoxide, and 7-deaza-dGTP—was essential to achieve amplification of DNA sequences of three disease genes showing a GC content ranging from 67 to 79%.