The polymerase chain reaction (PCR) is a powerful tool for the amplification of trace amounts of nucleic acids, and has rapidly become an essential analytical tool for virtually all aspects of biological research in experimental biology and medicine. Because the application of this tech ...

Historically, clinicians and researchers have relied upon the development of clinical endpoints or the use of surrogate markers in the evaluation of disease pathogenesis and in response to various therapeutic agents. In addition, microbiologic methods of detecting various path ...

Although reverse transcriptase polymerase chain reaction (RT-PCR) is an extremely sensitive method of mRNA analysis, obtaining quantitative information with this technique can be difficult. This is caused primarily by the fact that there are two sequential enzymatic steps invol ...

Quantitative PCR can be done either by measuring the amount of PCR products at a given number of cycle (end-point quantitative PCR) (1–4) or by following the amount of products during the PCR at several cycles (kinetic quantitative PCR) (5,6). In this chapter, we define these two quantitative PCR meth ...

Among the numerous assays proposed for quantifying specific nucleic-acid sequences in biological samples, PCR offers the greatest sensitivity and versatility. The assay for quantifying the amount of polymerase chain reaction (PCR) products is a crucial step in any quantitative PCR ...

The exponential amplification of small amounts of nucleic acids makes polymerase chain reaction (PCR) not only powerful but also challenging as a quantitative method. Variations in nucleic acid preparation, thermal cyclic performance, the choice of the polymerase, and the amplifi ...

In microbiology, the polymerase chain reaction (PCR) has become an important tool for the analysis of clinical samples. For example, it led to the detection of Hepatitis-B Virus DNA (HBV-DNA) in patients with serological patterns not previously associated with active infection (1). PCR-ba ...

Quantitative mRNA characterization by reverse transcription (RT) of RNA and subsequent polymerase chain reaction (PCR) (RT-PCR) is, compared to qualitative RT-PCR detection of RNA, more complicated because of two features inherent in in vitro amplification. First, during the expo ...

Detection of the products of the PCR reaction using nonisotopically labeled DNA molecules containing biotin, fluorescein, or digoxigenin has become a popular method for identification of specific products of polymerase chain reaction (PCR) (1,3). These labeled molecules are prep ...

Over the last nine years, several quantitative polymerase chain reaction (QPCR) techniques have been developed, and these are now frequently used for the quantification of DNA template copy numbers. However, only few of these PCR techniques are suitable for the precise and absolute quant ...

The PCR technique provides highly specific and sensitive means for analyzing nucleic acids, but it does not allow their direct quantification. This limitation originates from the fact that the efficiency of PCR depends on the amount of template sequence present in the sample, and the amplif ...

Many different protocols are now available for competitive polymerase chain reaction (PCR) and most rely on the use of a mimic or competitor that serves as a reference for quantitation (1–4). The success (or failure) of all these protocols is critically dependent on the design, construction, and ...

A real-time kinetic tool for polymerase chain reaction (PCR) quantitation, AmpliSensor assay (1) quantifies PCR product by relating the rate of an amplification reaction through the progressive depletion of a rate-limiting primer. AmpliSensor assay invokes a two-step amplifica ...

Reporter gene plasmids have been used extensively to monitor gene expression and elucidate intracellular pathways (1–4). They have been particularly useful in understanding the architecture of promoter regions and the interactions between promoter elements and cellular or vi ...

When studying the effect of various treatments on gene expression in humans, one occasionally is faced with the problem of detecting small changes in transcript levels in minute tissue samples. In addition, interindividual variations can be quite large and may even be the major source of vari ...

Quantitative polymerase chain reaction (PCR) is aimed to determine the absolute or relative amounts of RNA or DNA sequences in a given sample. There are two facts limiting the convenience of this approach. First, in most cases, only one or two sequences are amplified in a given round of amplification. ...

Polymerase chain reaction (PCR) is an important qualitative procedure in the routine microbiology laboratory for detecting the presence or absence of potentially harmful microorganisms in clinical specimens (1,2). The use of PCR to quantify an infectious agent in a clinical specimen ...

To assay gene expression or virus genomes in tissues or body fluids, competitive polymerase chain reaction (cPCR) is now performed in many laboratories. cPCR is a quantitative adaption of the PCR method in which a known number of copies of a synthetic RNA (1) or DNA (2–4) is coamplified with the target sam ...

The polymerase chain reaction (PCR) has revolutionized molecular biology. Portions of single-copy per cell genes (and cDNAs) prepared from very small tissue or cell samples can be specifically amplified for use in sequence determination, gene identification, and quantitation. Im ...

This brief guide is not intended as a full explanation of the theory and practice of nuclear magnetic resonance (NMR), on which there are a large number of excellent texts (1–3), but as an introduction to the terms used in the subsequent chapters. The section as a whole does not provide a comprehensive outli ...