[Time of flight mass spectrometry of DNA for rapid sequence determination]
The objective of this project is to develop a time-of-flight mass spectrometric approach to ordering Sanger sequence fragments, replacing electrophoresis and removing the electrophoresis bottleneck to rapid DNA sequencing, When the project was funded, we had demonstrated that massive DNA molecules could be volatilized, substantially intact, by a process involving pulsed laser ablation of a frozen film of a DNA solution. Using a crude time-of-flight mass spectrometer, we had demonstrated that ions of the ablated DNA could be formed, and that mass spectra were obtainable which appeared to contain only the parent molecular ion. The laser used was a dye laser which we tuned to match sodium atom resonances to increase the ionization efficiency. By pulsed laser ablation of frozen aqueous DNA solution films we have produced mass spectra of DNA mixtures which largely fulfil the simple requirement for DNA mixture analysis: one peak per DNA segment The peaks are clean and free of the fragment or adduct tails which characteristically degrade mass spectra obtained by UV laser ablation using UV chromophore matrices. To date, our approach has been characterized by extremely poor reproducibility; however the high quality of the mass spectra suggest that when better control of the ionization process is achieved, the use of an aqueous matrix offers an extremely promising approach to time-of-flight mass spectrometric sorting of DNA sequence mixtures.
- Research Organization:
- Arizona State Univ., Tempe, AZ (United States)
- Sponsoring Organization:
- USDOE; USDOE, Washington, DC (United States)
- DOE Contract Number:
- FG02-91ER61127
- OSTI ID:
- 7120637
- Report Number(s):
- DOE/ER/61127-2; ON: DE93006754
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
OLIGONUCLEOTIDES
MASS SPECTROSCOPY
TIME-OF-FLIGHT MASS SPECTROMETERS
PERFORMANCE TESTING
DNA SEQUENCING
PROGRESS REPORT
DOCUMENT TYPES
DYNAMIC MASS SPECTROMETERS
MASS SPECTROMETERS
MEASURING INSTRUMENTS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
SPECTROMETERS
SPECTROSCOPY
STRUCTURAL CHEMICAL ANALYSIS
TESTING
TIME-OF-FLIGHT SPECTROMETERS
550200* - Biochemistry
550400 - Genetics