Quantitative RT-PCR for Neuroendocrine Studies

Pedro L. Zamorano, Virendra B. Mahesh, Darrell W. Brann

Research output: Contribution to journalArticlepeer-review

51 Scopus citations

Abstract

Determination of mRNA levels of specific genes is becoming increasingly important as a measure of gene expression. With the recent advent of RT-PCR, the sensitivity for mRNA determination has been increased dramatically, and this technique is becoming widely used in neuroendocrine studies which involve small tissue samples and/or isolated nuclei. Nevertheless, the exact procedure for reliable quantification of RT-PCR has been widely debated. This minireview attempts to assimilate the available literature on the RT-PCR technique and discuss the various approaches commonly used to obtain quantitative results using the technique. An example from our laboratory of the use of RT-PCR for the measurement of several gene products in the same sample using exogenous internal standards is also provided. Particular attention is paid to the choice of endogenous vs. exogenous internal standards, the length of the transcript of the standard and its relationship to the target sequence being amplified, the amplification pattern of the target gene and internal standard, the reproducibility of the method, and the overall usefulness and suitability of RT-PCR for neuroendocrine studies.

Original languageEnglish (US)
Pages (from-to)397-407
Number of pages11
JournalNeuroendocrinology
Volume63
Issue number5
DOIs
StatePublished - 1996

Keywords

  • Molecular neuroendocrinology
  • Polymerase chain reaction

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Endocrinology
  • Endocrine and Autonomic Systems
  • Cellular and Molecular Neuroscience

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