PCR-RFLP and Real-Time PCR Techniques in Molecular Cancer Investigations

Abstract

The polymerase chain reaction (PCR) is a rapid scientific method for generating a 106-107fold increase in the number of copies of discrete DNA or RNA sequences (Boehm,1989; Imboden et al,1993). The use of PCR technology has greatly increased the ability to study on genetic material. PCR is a rapid and reliable molecular biology technique that allows quick replication of mainly DNA, the starting material can be a single molecule of rRNA or mRNA. It was developed by Kary Mullis in 1983, and he was awarded the Nobel Prize in 1993. PCR method is useful in situations of limited amount of DNA sample as in forensics, prenatal testing, because it amplifies a single or a few copies of DNA creating millions of copies of the region(1). The ability to quickly produce large quantities of genetic material has enabled significant scientific advances including DNA fingerprinting and sequencing of the human genome. As PCR technology allows taking specimen of genetic material even from just one cell, copy its genetic material several times, this facilitates genetic studies. Currently, besides research purposes, PCR technology is heavily used in diagnosis and patient management especially for viral diseases such as AIDS and hepatitis. Other than detection of infectious organisms, this technology is also useful for determination of genetic polymorphisms or mutations of individuals (Stahlberg,2011).