Quantitation of mRNA by Polymerase Chain Reaction
Nonradioactive PCR Methods
Published on 29. December 2011
Book
Paperback/Softback
XX, 166 pages
978-3-642-79714-9 (ISBN)
Description
In this laboratory "cook-book", the authors provide a concise guide to PCR-based techniques to quantify nucleic acids in biological and clinical samples using exclusively nonradioactive detection methods, e.g. HPLC, biotin and digoxigenin based protocols. Each method presentation also includes sections on theory, reagents, standards, applicability, limitations, and trouble shooting. In addition to the protocols, the authors also provide the necessary information on: general aspects of nucleic acid quantitation; design of PCR standards; mRNA purification; cDNA synthesis; solution hybridization; DNA sequencing. This laboratory guide enables professionals as well as beginners to adopt easily quantitative PCR protocols into their own clinical or biomedical research.
Reviews / Votes
"...this technical book will be very useful for all customers of PCR. And they are numerous!... Summarizing: an excellent booklet, which we recommend highly and absolutely necessary to all practicians of PCR products and their applications." Cellular and Molecular BiologyMore details
Series
Edition
Softcover reprint of the original 1st ed. 1995
Language
English
Place of publication
Berlin
Germany
Publishing group
Springer Berlin
Target group
Professional and scholarly
Research
Illustrations
17 s/w Abbildungen
XX, 166 p. 17 illus.
Dimensions
Height: 235 mm
Width: 155 mm
Thickness: 11 mm
Weight
306 gr
ISBN-13
978-3-642-79714-9 (9783642797149)
DOI
10.1007/978-3-642-79712-5
Schweitzer Classification
Other editions
Additional editions
Thomas Köhler | Dirk Laßner | Anne-Katrin Rost
Quantitation of mRNA by Polymerase Chain Reaction
Nonradioactive PCR Methods
Book
10/1995
Springer
€85.55
Article exhausted; check different version
Content
I Theoretical and Methodical Prerequisites for Using PCR to Quantitate Nucleic Acids.- 1.1 General Aspects and Chances of Nucleic Acid Quantitation by PCR.- 1.2 Design of Suitable Primers and Competitor Fragments for Quantitative PCR.- 1.3 Cloning of Short DNA Fragments and In Vitro Transcription to Generate RNA Standards.- 1.4 Direct Non-lsotopic Sequencing of PCR Products or Standards.- 2Conventional Techniques for mRNA Analysis.- 2.1 Isolation of mRNA.- 2.2 Synthesis of cDNA.- 2.3 Qualitative RT-PCR: Amplification of Synthesized mdr-1 cDNA.- 2.4 Single-Tube RT-PCR.- 2.5 Nonradioactive Determination of PCR Products by Using a DIG-Labeled DNA Probe (Dot Blot).- 2.6 Nonradioactive Northern Blot Hybridization with DIG-Labeled DNA Probes.- 3 Semiquantitative and Quantitative Protocols for Measurement of Nucleic Acids by PCR.- 3.1 Quantitation of mRNA by the ELOSA Technique Using External Standards.- 3.2 Semiquantitative Detection of Viral DNA, e.g. for CMV, by Using the DNA Enzyme Immunoassay (DEIA).- 3.3 HPLC-Analysis of Nucleic Acids.- 3.4 Quantitation of Absolute Numbers of mRNA Copies in a cDNA Sample by Competitive PCR.- Acknowledgment.