Quantitative PCR. A survey of the present technology

Area of Science:

• Molecular Biology
• Biochemistry
• Clinical Diagnostics

Background:

• Polymerase Chain Reaction (PCR) is a fundamental technique in molecular biology and clinical diagnostics.
• Quantitative data is often required to correlate amplified nucleic acid product with the initial target amount.
• The exponential nature of PCR allows for adaptation to quantitative applications.

Purpose of the Study:

• To present various strategies for sensitive and specific detection and quantification of PCR products.
• To discuss the statistical aspects, advantages, and limitations of different quantitative PCR protocols.
• To outline the suitability of conventional and advanced techniques for precise amplification product determination.

Main Methods:

• Review of basic strategies including external and internal standards for PCR quantification.
• Discussion of conventional laboratory techniques: gel electrophoresis and High-Performance Liquid Chromatography (HPLC).
• Exploration of non-radioactive labeling procedures and solid-phase-mediated strategies for product detection.

Main Results:

• Various methods for sensitive and specific PCR product detection and quantification are presented.
• Statistical aspects, advantages, and limitations of different quantitative PCR protocols are discussed.
• Suitability of conventional and advanced techniques for precise determination of amplification products is outlined.

Conclusions:

• Quantitative PCR offers powerful tools for measuring nucleic acids in diverse applications.
• A range of strategies exists for sensitive and specific PCR product quantification, each with unique benefits and drawbacks.
• Selection of appropriate methods, including conventional and advanced techniques, is crucial for accurate nucleic acid quantification.