Nested Pcr Protocol
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RT-PCR Protocols
Author: Nicola King
language: en
Publisher: Springer Science & Business Media
Release Date: 2008-02-04
Until the mid 1980s, the detection and quantification of a specific mRNA was a difficult task, usually only undertaken by a skilled molecular biologist. With the advent of PCR, it became possible to amplify specific mRNA, after first converting the mRNA to cDNA via reverse transcriptase. The arrival of this technique—termed reverse transcription-PCR (RT-PCR)—meant that mRNA suddenly became amenable to rapid and sensitive analysis, without the need for advanced training in molecular biology. This new accessibility of mRNA, which has been facilitated by the rapid accumulation of sequence data for human mRNAs, means that every biomedical researcher can now include measurement of specific mRNA expression as a routine component of his/her research plans. In view of the ubiquity of the use of standard RT-PCR, the main objective of RT-PCR Protocols is essentially to provide novel, useful applications of RT-PCR. These include some useful adaptations and applications that could be relevant to the wider research community who are already familiar with the basic RT-PCR protocol. For example, a variety of different adaptations are described that have been employed to obtain quantitative data from RT-PCR. Quantitative RT-PCR provides the ability to accurately measure changes/imb- ances in specific mRNA expression between normal and diseased tissues.
PCR Protocols
Author: John M.S. Bartlett
language: en
Publisher: Springer Science & Business Media
Release Date: 2003-08
Drawing on the proven qualities of the much praised and widely used first edition, John M. S. Bartlett and David Stirling have thoroughly updated and dramatically expanded the number of protocols to take advantage of the newest technologies used in all branches of research and clinical medicine today. These successful methods include real-time PCR, SNP analysis, nested PCR, direct PCR, and long-range PCR. Among the highlights are chapters on genome profiling by SAGE, differential display and chip technologies, the amplification of whole genome DNA by random degenerate oligonucleotide PCR, and the refinement of PCR methods for the analysis of fragmented DNA from fixed tissues. In situ PCR methods and their application in parallel with other methods, such as immunohistochemistry, are also included. Each fully tested protocol is described in step-by-step detail by an established expert in the field and includes a background introduction outlining the principle behind the technique, equipment and reagent lists, tips on troubleshooting and avoiding known pitfalls, and, where needed, a discussion of the interpretation and use of results. Cutting-edge and highly practical, PCR Protocols, Second Edition provides both novice and experienced investigators with an up-to-date compendium of powerful PCR methods for easy reference and consultation in the day-to-day performance of PCR-based experimentation, one that will enhance understanding of PCR, satisfy current needs, and point to powerful future applications.
Principles and Technical Aspects of PCR Amplification
Author: Elizabeth van Pelt-Verkuil
language: en
Publisher: Springer Science & Business Media
Release Date: 2008-03-14
Kary Mullis was awarded a Nobel Prize for inventing the PCR technique more than a decade ago in 1993. Since its "discovery", multiple adaptations and variations of the standard PCR technique have been described. This publication aims to provide the reader with a guide to the standard PCR technique and its many available variants, with particular emphasis being placed on the role of these PCR techniques in the clinical diagnostic laboratory (the central theme of this book).