细胞功能测定

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 ...

Nuclear magnetic resonance (NMR) spectroscopy has become established in recent years as a uniquely powerful technique for studying the structures of proteins in solution. In a 1H spectrum, each hydrogen atom in the molecule gives rise to an individual signal, and in favorable cases, it is poss ...

Developing nerve growth factor (NGF)-dependent sympathetic neurons are one of the best-studied in vitro models of neuronal apoptosis and have been used to identify key components of the neuronal cell death pathway. This chapter describes how to prepare purified cultures of primary symp ...

Mitogen-activated protein kinases (MAPKs) are activated by a wide variety of cellular stimuli and involved in the regulation of most, if not all, cellular processes. Among them, the c-Jun N-terminal kinase and p38 MAPKs are predominantly induced in response to proinflammatory cytokines a ...

Developing sympathetic neurons, which depend on nerve growth factor for survival, are one of the best studied in vitro models of neuronal apoptosis and have been extensively used for cellular and molecular studies of the neuronal death pathway. Important apoptotic events after nerve gro ...