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๐Ÿ“

Modern Applications of DNA Amplification Techniques: Problems and New Tools

โœ Scribed by Tanya Vener, Malin Stark, Jan Albert, Mathias Uhlรฉn, Joakim Lundeberg (auth.), Dirk Lassner, Barbara Pustowoit, Arndt Rolfs (eds.)


Publisher
Springer US
Year
1997
Tongue
English
Leaves
143
Edition
1
Category
Library

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โœฆ Synopsis


In the ten years since the first publication on PCR (Saiki et al. , 1985), this in vitro method of nucleic acid replication and modification has grown to rival in popularity traditional microbiological, genetical und technical procedures for cloning, sequencing, gene detecting and related procedures. To date the PCR literature has emphasized six main areas of application: genetic mapping, detection of mutations, genetic polymorphism, transcriptional splicing and regulation, molecular virology and quantitative procedures. The overwhelming focus of quantification of DNA or RNA by PCR has been on human microbiology and oncological problems. The exquisite sensitivity of PCR gives this method the ability to detect extremely rare DNAs, mRNAs, mRNAs in small numbers of cells or in small amounts of tissue, and mRNAs expressed in mixed-cell populations. However, the exact and accurate quantification of specific nucleic acids in biological samples is in spite of numerous publications in that field still a general problem: during the peR process, an unknown initial number of target sequences are used as a template from which a large quantity of specific product can be obtained. Although the amount of product formed is easy to determine, it is difficult to deduce the initial copy number of the target molecule because the efficiency of the peR is largely unknown.

โœฆ Table of Contents


Front Matter....Pages I-VIII
Front Matter....Pages 1-1
Multiple Competitors for Single-Tube Quantification of HIV-1 DNA....Pages 3-10
Quantitation of P53 Tumor Suppressor Gene Copy Number in Tumor Dna Samples by Competitive PCR in an Elisa-Format....Pages 11-23
Standardisation of Messenger RNA Quantification Using an RT-PCR Method Involving Co-Amplification with a Multi-Specific Internal Control....Pages 25-35
Quantitative Analysis of Human DNA Sequences by Solid-Phase Minisequencing....Pages 37-45
Bioimage Analysers: Application for Ribozyme Kinetics....Pages 47-54
First Approaches to Quantitate MDR1-Messenger RNA by In Cell PCR....Pages 55-63
Application of In Situ-PCR for the Detection of Intracellular mRNAs....Pages 65-76
Psoralen Biotin: A Novel Reagent for Non-Enzymatic and Specific Labeling of Nucleic Acid Probes and Oligonucleotides....Pages 77-82
The Effect of Quantitative Ratio Between Primer Pairs on Pcr Products in Multi-Target Amplification....Pages 83-89
Front Matter....Pages 91-91
Quantitation of Rubella Virus Genome by QPCR and Its Application to Resolution for Mechanism of Congenital Rubella Syndrome....Pages 93-100
Significance of the Detection of Rubella Virus Rna by Nested PCR in Prenatal Diagnostics of Viral Infections....Pages 101-108
Quantitative Detection of Human Cytomegalovirus DNA in Cerebrospinal Fluid by Polymerase Chain Reaction....Pages 109-117
Quality Control and External Quality Assessment Schemes for the Diagnostic Use of PCR In Microbiological Laboratories โ€” European Trials on Hepatitis B Virus and Cytomegalovirus....Pages 119-128
Back Matter....Pages 129-143

โœฆ Subjects


Animal Anatomy / Morphology / Histology; Biotechnology


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