Expression profiling of tubulin isotypes and microtubule-interacting proteins using real-time polymerase chain
Sharon Lobert1, Laree Hiser, John J Correia
1School of Nursing, University of Mississippi Medical Center, Jackson, Mississippi 39216, USA.
Abstract:
Real-time polymerase chain reaction (PCR) has been used for quantification of intracellular mRNA levels in cell culture and tissue samples. It is an important tool for studying antimitotic drug effects on tubulin isotype and microtubule-interacting protein levels and for measuring differences in normal and tumor tissue samples that could have predictive or prognostic applications. Both quantitative and comparative methods are valuable approaches; however, the selection of either approach requires an understanding of their benefits and challenges. In this chapter, we provide detailed protocols for real-time PCR experiments, discuss issues to consider in selecting real-time PCR methodologies, and give examples utilizing either quantitative or comparative approaches.
Insights
Real-time polymerase chain reaction (PCR) quantifies mRNA in cells and tissues. This guide details PCR methods for studying drug effects and cancer biomarkers, aiding researchers in choosing the best approach.
Area of Science:
- Molecular Biology
- Biochemistry
- Genomics
Background:
- Real-time polymerase chain reaction (PCR) is crucial for quantifying intracellular mRNA levels.
- It aids in studying antimitotic drug effects on tubulin and microtubule-interacting proteins.
- PCR is valuable for analyzing normal versus tumor tissue for prognostic and predictive insights.
Purpose of the Study:
- To provide detailed protocols for real-time PCR experiments.
- To discuss considerations for selecting appropriate real-time PCR methodologies.
- To illustrate quantitative and comparative PCR approaches with examples.
Main Methods:
- Detailed protocols for real-time PCR experiments are presented.
- Methodologies for quantitative and comparative real-time PCR are discussed.
- Examples of both quantitative and comparative approaches are provided.
Main Results:
- The chapter offers a comprehensive guide to performing real-time PCR.
- It clarifies the benefits and challenges of quantitative versus comparative methods.
- Practical examples demonstrate the application of these techniques.
Conclusions:
- Understanding real-time PCR methodologies is essential for accurate mRNA quantification.
- The choice between quantitative and comparative approaches depends on experimental goals.
- This chapter equips researchers with the knowledge to effectively apply real-time PCR.
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