Routes to improving the reliability of low level DNA analysis using real-time PCR.

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.

Results

At very low target concentrations of 0.5-10 genome equivalents (g.e.) eliminating any replicates within each DNA standard

SUBMITTER: Ellison SL 

PROVIDER: S-EPMC1559608 | biostudies-literature | 2006 Jul

REPOSITORIES: biostudies-literature