Routes to improving the reliability of low level DNA analysis using real-time PCR |
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Authors: | Stephen LR Ellison Claire A English Malcolm J Burns Jacquie T Keer |
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Institution: | (1) Analytical Technology, LGC Limited, Teddington, TW11 0LY, UK |
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Abstract: | Background Accurate quantification of DNA using quantitative real-time PCR at low levels is increasingly important for clinical, environmental
and forensic applications. At low concentration levels (here referring to under 100 target copies) DNA quantification is sensitive
to losses during preparation, and suffers from appreciable valid non-detection rates for sampling reasons. This paper reports
studies on a real-time quantitative PCR assay targeting a region of the human SRY gene over a concentration range of 0.5 to
1000 target copies. The effects of different sample preparation and calibration methods on quantitative accuracy were investigated. |
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