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A procedure for analysis of densitometric spectra
1Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA.
Electrophoresis
|October 1, 1989
Summary
Accurate computer-aided analysis of densitometric spectra requires careful selection of component band shapes. This method enhances quantitative analysis for DNA and gel electrophoresis data.
Area of Science:
- Biophysics
- Analytical Chemistry
- Computational Biology
Background:
- Densitometric spectra analysis is crucial for quantifying biological and chemical samples.
- Accurate decomposition of overlapping spectral bands presents a significant analytical challenge.
Purpose of the Study:
- To develop and validate a computer-aided method for accurate quantitative analysis of densitometric spectra.
- To assess the impact of different component band shapes on spectral decomposition accuracy.
Main Methods:
- Utilized Powell and Marquardt minimizers for decomposing densitometric spectra into component bands.
- Evaluated various functions (Cauchy, Gauss, cosine) for approximating band shapes.
- Applied the method to analyze DNA cleaved by neocarzinostatin.
Main Results:
- High accuracy in spectral decomposition is achievable with appropriately chosen band shapes.
- Asymmetric Cauchy and Gauss functions effectively approximate highly overlapped bands.
- The cosine function provides high accuracy for asymmetric densitometric curves.
Conclusions:
- The developed method offers a general approach for quantitative analysis of densitometric data.
- Accurate spectral fitting is dependent on selecting suitable mathematical functions for band shapes.
- This technique is applicable to autoradiographic films and electrophoretic gel scans.