ABSTRACT
This chapter reviews the nucleic acid amplification approaches, providing not only the theory behind how they work, but also detailed explanations of various experimental applications. One approach to nucleic acid amplification utilizes both in situ transcription and anti-sense ribonucleic acid amplification in order to obtain abundant yields of desired targets. The polymerase chain reaction (PCR) is a powerful amplification technique that has revolutionized the study of gene expression in both normal and abnormal tissues. The three steps of conventional PCR amplification are: denaturation of the template deoxyribonucleic acids (DNA), annealing of the oligonucleotide primers, and cDNA synthesis. Equipment used for PCR should be kept in closed containers and away from possible contamination by exogenous DNA. PCR amplifications often result in indeterminate results or multiple cDNA products. Some of these cDNAs are artifactual or represent false-positives due to exogenous DNA contamination. The PCR has direct applications in the area of differential gene expression.
