Quantitation of mRNA by Polymerase Chain Reaction Nonradioactive PCR Methods
In this laboratory "cook-book", the authors provide a concise guide to PCR-based techniques for quantifying nucleic acids in biological and clincial samples using exclusively nonradioactive detection methods, e.g. HPLC, biotin and digoxigenin based protocols. Each method presentation also...
| Main Authors: | , , , |
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| Format: | eBook |
| Language: | English |
| Published: |
Berlin, Heidelberg
Springer Berlin Heidelberg
1995, 1995
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| Edition: | 1st ed. 1995 |
| Series: | Springer Lab Manuals
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| Subjects: | |
| Online Access: | |
| Collection: | Springer Book Archives -2004 - Collection details see MPG.ReNa |
Table of Contents:
- 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