Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1994-09-08
1997-02-18
Chambers, Jasemine C.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
435 32, C12Q 168, C12Q 118
Patent
active
056040962
ABSTRACT:
The present invention pertains to methods of monitoring mycoplasmal growth whereby a culture of mycoplasmal cells is contacted with a fluorescent nucleic acid-binding agent under conditions which allow the agent to selectively bind to mycoplasmal nucleic acid. The amount of fluorescence associated with the mycoplasmal nucleic acid indicates the number of mycoplasmal cells in the culture. The fluorescence of the culture is typically read using a fluorometer which correlates with colonial growth on agar. A particularly preferred fluorescent nucleic acid-binding agent is ethidium bromide. The present invention further pertains to methods of determining the effectiveness of an antimycoplasmal antibiotic.
REFERENCES:
patent: 2662082 (1953-12-01), Short et al.
Valle et al. (1981) "Fluorometric Determination of DNA & RNA in Chlamydamonas using Ethidium Bromide" J Biochem Biophys Meth. 4:271-277.
Le Pecq et al. (1966) "A New Fluorometric Method for RNA & DNA Determination" Anal. Biochem. 17: 100-107.
Halprin et al. (1979) A combined Alkali extraction-Ethidium Bromide Technique for the Measurement of DNA in Small Pieces & Tissue. J. Invest. Dermat. 73:359-363.
LePecq et al. (1967) "A Fluorescent Complex between Ethidium Bromide and Nucleic Acids" J. Mol. Biol 27:87-106.
Sambrook et al. (1989) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press. Appendix E5.
Latt et al., "Fluorescent Probes of DNA Microstructure and DNA Synthesis" Flow Cytometry and Sorting, Second Edition pp. 249-290 (1990).
Kowalski "A Procedure for the Quantitation of Relaxed Closed Circular DNA in the Presence of Superhelical DNA: An Improved Fluorometric Assay for Nicking-Closing Enzyme" Analytical Biochemistry 1979, vol. 93, pp. 346-354 .
Melamede Robert
Schaeffer Warren I.
Chambers Jasemine C.
Priebe Scott D.
Remillard Jane E.
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