Comparison of different baterial genomic dna extraction protocol selected gram positive bacteria

Amelia, Ausgistine Te-eng. (2006) Comparison of different baterial genomic dna extraction protocol selected gram positive bacteria. [Final Year Project Report] (Unpublished)

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Abstract

In this study, four different DNA extraction protocols were perfonned on three different Gram positive bacterial species to determine the best protocols based on the DNA quantity and purity of the extracted DNA. The evaluation on the DNA quantity and purity were determined by using spectrophotometer and two PCR methods which were Specific PCR and ERIC-PCR. Two species used were from the genus Listeria and one species was from the genus Staphylococcus. The organisms were L. monocytogenes (ATCC 7644), L. innocua (ATCC 33090) and S. aureus (ATCC 25923). For the extraction process, three conventional methods were perfonned on the selected bacterial species namely cell boiling, PCI and CTAB and Promega Wizard® Genomic Purification System (Promega, USA). Serial dilution was done for up to 10-5 dilution. The sensitivity of the methods was also compared between the different number of bacterial count. The overall results of the study indicated that the commercial extraction kit provided satisfying results in the PCR analysis. The most promising method based on DNA quantity and purity differ for different bacterial species.

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