Type

Journal Article

Authors

M Clynes
M Saleh
C Daly
L O'Driscoll

Subjects

Microbiology

Topics
specific reverse transcriptase polymerase chain reaction polymerase chain reaction carboxylic ester hydrolases carrier proteins beta 2 microglobulin humans gene expression pcr cells in situ hybridization artifacts p glycoprotein genetics metabolism carboxylesterase biosynthesis rna directed dna polymerase membrane glycoproteins indicators and reagents esd protein human drug resistance animals actins rna messenger isolation purification dna primers

The use of reverse transcriptase-polymerase chain reaction (RT-PCR) to investigate specific gene expression in multidrug-resistant cells. (1993)

Abstract Expression of specific genes at the level of mRNA can be studied using techniques such as Northern blot, slot/dot blot, RNase protection assay, 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-PCR technique is described in detail with sections dealing with RNA extraction, choice of primers (including the use of cDNA sequence data bases), PCR and RT-PCR protocols in addition to the limitations of the method. The study of one particular mRNA transcript (MDR1) using RT-PCR is discussed in detail. Recently described methods for quantitation of PCR products are discussed. Quantitative PCR would appear to offer a method of studying gene expression in a more extensive way than has been possible to date.
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Full list of authors on original publication

M Clynes, M Saleh, C Daly, L O'Driscoll

Experts in our system

1
Martin Clynes
Dublin City University
Total Publications: 209
 
2
C Daly
Trinity College Dublin
Total Publications: 15
 
3
Lorraine O'Driscoll
Trinity College Dublin
Total Publications: 152