The Use of Real-Time Reverse Transcription-PCR for Assessing Estrogen Receptor and Estrogen-Responsive Gene Expression

Part of the Methods in Molecular Biology book series (MIMB, volume 1366)

Abstract

Real-time reverse transcription-polymerase chain reaction (RT-PCR), also known as quantitative RT-PCR (qRT-PCR), is a powerful tool for assessing gene transcription levels. The technique is especially useful for measuring estrogen receptor transcript levels as well as gene expression changes in response to estrogen stimulation as it is quick, accurate, robust, and allows the measurement of gene expression in a variety of tissues and cells. This chapter describes the protocols used for the real-time RT-PCR assay using hydrolysis (TaqMan-type) probes.

Key words

Estrogenreceptors Gene expression Primer Fluorogenic probe Real-time RT-PCR Master mix Custom design Normalization 

Notes

Acknowledgements

This work was supported, in part, by NIH P20GM103443. The authors would like to thank Dr. Rozzy Finn for reviewing this chapter.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Sanford ResearchSioux FallsUSA
  2. 2.Division of Basic Biomedical Sciences, Sanford School of MedicineUniversity of South DakotaVermillionUSA

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