An Efficient Method for DNA Isolation and Purification from Vibrio fluvialis | ||
| Al-Mustansiriyah Journal of Science | ||
| Article 1, Volume 23, Issue 5, July 2012, Pages 25-32 | ||
| Author | ||
| Iyad J. Qassar | ||
| Abstract | ||
| An efficient procedure for the isolation and purification of total DNA has been developed for the gram-negative bacterium Vibrio fluvialis, which was found to be difficult to be isolated using the published methods. According to this procedure the cell suspension was incubated with an enzyme mixture containing lysozyme, RNase, and proteinase K, allowing each enzyme to work independently of the other by varying the incubation temperature. Therefore, shearing and degradation of the DNA as a result of extended manipulation and possible inadequate handling of the lysate was avoided. The average DNA yields obtained were in the range of 1mg per 10ml of cell suspension, with a high level of purity of about 1.8-1.9. Examination of the DNA preparations using agarose gel electrophoresis revealed a well defined single band free of contaminating RNA and residual DNA. Finally, this DNA was found to be a good substrate for restriction enzymes tested and ligation experiments. | ||
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