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Food Control, 34, 1, 47 - 49
Biochem Biophys Res Commun, 390, 3, 733-737
Genet Test Mol Biomarkers 19,2,63-68
Aims: The aim of this study was to establish an effective real-time quantitative polymerase chain reaction (qPCR) method for telomere length measurement on the Roche LightCycler® 480 (LC480) real-time PCR platform.
Methods: Measurement of relative average telomere length was achieved by comparing products amplified from telomere-specific primers and single copy reference gene primers in a ratio (T/S).
Results: Extensive testing led us to conclude that a modification of the original two-plate T/S assay was more compatible with this platform than the recently developed single-plate assay, and that choice of hot-start Taq polymerase and intercalating dye were critical factors.
Conclusions: This modified assay generates reliable measurements as judged by correlation with data derived by the telomeric restriction fragment Southern blot-based method.
Specific Amplification of Difficult PCR Products from Small Amounts of DNA Using FastStart Taq DNA Polymerase
High Pure PCR Cleanup Micro Kit
Add more flexibility to nucleic acid purification - quickly and efficiently purify products from PCR and other reactions.
Mutation Detection Using Multi-color Detection on the LightCycler System
The LightCycler System is a powerful tool for amplifying and quantitating PCR products in a very short time (Wittwer et al. 1997). By using melting curve analysis, the system also provides a unique and innovative approach for the detection of single nucleotide polymorphisms.
Quantitative PCR by Continuous Fluorescence Monitoring of a Double Strand DNA-Specific Binding Dye
A simple method for quantitative PCR using the LightCyclerTM and the dsDNA binding dye SYBR® Green I Dye is demonstrated. The method has a dynamic range of at least six orders of magnitude and a sensitivity of a single copy per reaction. Two methods for removing the confounding signal from nonspecific amplification products are described. The method is suitable for both DNA and RNA quantification.
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