THE USE OF REVERSE TRANSCRIPTASE-POLYMERASE CHAIN-REACTION (RT-PCR) TO INVESTIGATE SPECIFIC GENE-EXPRESSION IN MULTIDRUG-RESISTANT CELLS

Expression of specific genes at the level of mRNA can be studied using techniques such as Northern blot, slot/dot blot, RNase protection ass ay, in situ hybridisation and RT-PCR. In this article these methods of analysis are compared; RT-PCR offers higher levels of specificity and sensitivity than traditional methods of RNA analysis and as such has become the method of choice for the study of gene expression. The RT-P CR technique is described in detail with sections dealing with RNA ext raction, choice of primers (including the use of cDNA sequence data ba ses), PCR and RT-PCR protocols in addition to the limitations of the m ethod. The study of one particular mRNA transcript (MDR1) using RT-PCR is discussed in detail. Recently described methods for quantitation o f PCR products are discussed. Quantitative PCR would appear to offer a method of studying gene expression in a more extensive way than has b een possible to date.