COMPARISON OF THREE DIFFERENT METHODS OF GENOMIC DNA EXTRACTION FROM GRAM POSITIVE AND GRAM NEGATIVE BACTERIA

Author (s): M. K. Hossain Ripon, M. Hasan, M. M. Ahasan*, M. W. Alam and Shaila Kabir

*Corresponding author, E-mail: labu63@gmail.com

 Abstract

Direct extraction of DNA from gram positive and gram negative microorganisms is still now challenging. In this study, a comparison was made between the relative efficacy of extraction of bacterial DNA from both gram negative (Pseudomonas aureus, E.coli, Shigella bodyii) and gram positive origin (Bacillus subtilis, Staphylococcus aureus, Bacillus cereus) from pure culture using three extraction methods. For all the gram positive and gram negative strains the DNA extracted by Boiling and Phenol chloroform were found to have almost similar value of DNA concentration than Bead beating methods. Extracted DNA concentrations found by Phenol chloroform method were 320, 255, 185, 350, 265, 295 g ml-1 respectively from Bacillus subtilis (+) Staphylococcus aureus (+) Bacillus cereus (+), Pseudomonas aureus (-), E.coli (-), Shigella bodyii (-). The extracted DNA concentrations by Boiling method were 350, 300, 450, 275, 310, 115 g ml
-1 respectively from same strains. The strains had the extracted DNA concentrations in Bead beating method of 330, 45, 90, 265, 125 and 50 g ml -1 respectively from same strains. The results indicated that Phenol chloroform and Boiling methods might be a suitable choice for extracting DNA from gram positive and gram negative strains. For Bead beating method, high molecular weight intact DNA bands were not observed but when extraction process were carried out by Phenol chloroform and Boiling methods, high molecular weight intact bands were observed in both the gram positive and gram negative microbes.

Key words: Extraction, Genomic DNA, Gram negative bacteria, Gram positive bacteria

Full  PDF: Click here

J. Expt. Biosci. 2(1): January 2011


data:newerPageTitle data:olderPageTitle data:homeMsg