In Situ Polymerase Chain Reaction: Overview of Procedures and Applications

Abstract

The evaluation of gene expression in the context of cellular morphology is essential to the full understanding of cell biology. A variety of methods for detection of nucleic acids are currently available. Solution polymerase chain reaction (PCR) requires disruption of the sample and detection of the amplified material by electrophoresis in agarose gels. In situ hybridization methods, on the other hand, permit morphologic correlation and provide a high sensitivity sufficient for many applications. In some instances, however, the amount of target in the sample is below the limit of detection of this technique. In situ PCR allows the detection of minimal amounts of nucleic acids with exquisite sensitivity and specificity, while the integrity of the cells and the morphology of tissues remains preserved. This technique, although not exempt from difficulties, is undergoing methodologic simplifications that will make it suitable for an increasing number of basic science and clinical applications. The following is a review of the principles, methods, and applications of in situ PCR.

Key Words:

• In situ PCR
• methods