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Updated: Jul 30, 2026
06:15
Concentration Determination of Nucleic Acids and Proteins Using the Micro-volume Bio-spec Nano Spectrophotometer
Published on: February 17, 2011
Comparison of spectrometric and autophotographic methods for determining DNA concentrations
1Microbiology Laboratory, Indiana University School of Medicine-Evansville Center 47712.
Biochemical and Biophysical Research Communications
|September 15, 1994
Summary
This study introduces a non-destructive DNA quantitation method using lumigenic detection and digitized X-ray films. This approach improves upon traditional methods by preserving the membrane for future analysis.
Area of Science:
- Molecular Biology
- Biochemistry
- Biotechnology
Background:
- Traditional DNA quantitation methods often involve radioactive probes and destructive sample processing.
- Lumigenic detection offers an alternative, but cutting membrane spots for assay is time-consuming and consumes the sample.
- Previous work established lumigenic detection with liquid scintillation counters for DNA quantitation.
Purpose of the Study:
- To develop and validate a non-destructive DNA quantitation method using lumigenic detection and digitized X-ray films.
- To compare the efficacy of digitized X-ray film analysis with traditional transmittance measurements for DNA quantitation.
- To assess the reliability of image analysis for determining DNA concentrations while preserving the experimental membrane.
Main Methods:
- Utilized a lumigenic detection method with liquid scintillation counters for DNA quantitation.
- Compared transmittance measurements with counts per minute for target CAT DNA.
- Employed digitized X-ray films read by image analysis for DNA concentration determination.
- Assessed the ability to distinguish between different picogram concentrations of target DNA.
Main Results:
- Both transmittance and lumigenic methods distinguished between 10 and 100 picograms of target DNA.
- Neither method could significantly distinguish DNA amounts below 10 picograms.
- Digitized X-ray film analysis provided reliable DNA concentration determination.
- The membrane remained intact after analysis, serving as a permanent record.
Conclusions:
- Digitized X-ray film analysis of lumigenic signals offers a reliable and non-destructive method for DNA quantitation.
- This technique eliminates the need for radioactive probes and preserves the membrane for future use.
- The method provides a valuable alternative for DNA analysis in molecular biology research.
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