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A Robust Curve-Fitting Procedure for the Analysis of Plasmid DNA Strand Break Data from Gel Electrophoresis

DOI: 10.1667/RR2514.1 DOI Help
PMID: 21466384 PMID Help

Authors: Stephen Mcmahon (Queen's University, Belfast) , Fred Currell (Queen's University)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Radiation Research , VOL 175 (6) , PAGES 797-805

State: Published (Approved)
Published: April 2011

Abstract: A robust method for fitting to the results of gel electrophoresis assays of damage to plasmid DNA caused by radiation is presented. This method makes use of nonlinear regression to fit analytically derived dose–response curves to observations of the supercoiled, open circular and linear plasmid forms simultaneously, allowing for more accurate results than fitting to individual forms. Comparisons with a commonly used analysis method show that while there is a relatively small benefit between the methods for data sets with small errors, the parameters generated by this method remain much more closely distributed around the true value in the face of increasing measurement uncertainties. This allows for parameters to be specified with greater confidence, reflected in a reduction of errors on fitted parameters. On test data sets, fitted uncertainties were reduced by 30%, similar to the improvement that would be offered by moving from triplicate to fivefold repeats (assuming standard errors). This method has been implemented in a popular spreadsheet package and made available online to improve its accessibility.

Subject Areas: Biology and Bio-materials, Physics, Medicine

Instruments: B16-Test Beamline

Added On: 09/04/2011 21:04

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