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A Non-Radioactive Method for Inexpensive Quantitative RT-PCR
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M. Maggiolini
Veröffentlicht/Copyright:
1. Juni 2005
Abstract
We present a novel method for quantitative RT-PCR that involves direct incorporation of digoxigenin-11-dUTP (DIG-dUTP) during amplification of cDNAs, separation of RT-PCR products by agarose gel electrophoresis, Southern transfer to a nylon membrane, and chemiluminescent detection with an anti-DIG antibody. The whole procedure can be done in about a day and has the following advantages: It is highly sensitive, specificity is confirmed by monitoring the size of the RT-PCR product, it is non-radioactive, quantitative, and does not require expensive specialized equipment.
Published Online: 2005-6-1
Published in Print: 1999-6-1
Copyright © 1999 by Walter de Gruyter GmbH & Co. KG
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Artikel in diesem Heft
- To Our Authors, Readers and Subscribers
- Michael Strauss. January 12, 1950 April 29, 1999
- Monoallelic Expression of Pax5: A Paradigm for the Haploinsufficiency of Mammalian Pax Genes?
- Adeno-Associated Viral Vectors for Gene Transfer and Gene Therapy
- Histidyl-tRNA Synthetase
- Dequalinium TM Vesicles Form Stable Complexes with Plasmid DNA which Are Protected from DNase Attack
- On the Role of Thymopoietins in Cell Proliferation. Immunochemical Evidence for New Members of the Human Thymopoietin Family
- Functional Characterization of Atherosclerosis-Associated Ser128Arg and Leu554Phe E-Selectin Mutations
- Cycloheximide, a New Tool to Dissect Specific Steps in ER-Associated Degradation of Different Substrates
- Localization of Rat Cathepsin K in Osteoclasts and Resorption Pits: Inhibition of Bone Resorption and Cathepsin K-Activity by Peptidyl Vinyl Sulfones
- Oxidation of 4-tert-Butylcatechol and Dopamine by Hydrogen Peroxide Catalysed by Horseradisch Peroxidase
- A Non-Radioactive Method for Inexpensive Quantitative RT-PCR
- Activation of the Transcription Factor ISGF3 by Interferon-gamma
- Acquisition of Myogenic Specificity through Replacement of One Amino Acid of MASH-1 and Introduction of an Additional alpha -Helical Turn
- The Heavy Metal-Responsive Transcription Factor-1 (MTF-1) Is Not Required for Neural Differentiation
- Human Fc gamma Receptor IIb Expressed in <I>Escherichia coli</I> Reveals IgG Binding Capability
- Mutational Analysis of Two Stefin A Epitopes
