Journal of Pure and Applied MicrobiologyVol. 7 No. 4

Fast Protocol for DNA Isolation of DNA from Bacterial Isolated from a Hyper-Arid Environment

Vianey Mendez-Trujillo1, Lizbeth Moreno-Ram?rez2, Monica Carrillo-Beltran1 and Daniel Gonz?lez-Mendoza2

1Universidad Aut?noma de Baja California, Instituto de Ingenier?a, Blvd. B. Ju?rez s/n, Col. Insurgentes Este, 21280 Mexicali, Baja California, Mexico. 2Instituto de Ciencias Agr?colas, Universidad Aut?noma de Baja California (ICA-UABC), Carretera a Delta s/n Ejido Nuevo Le?n, 21705, Baja California, Mexico.

Received on 18 January 2013 and accepted on 26 February 2013

 

ABSTRACT

The extraction of high quality genomic DNA for PCR amplification from phosphate solubilising microbes (gram positive) is complicated due to the presence of a high percentage of secondary metabolites or PCR inhibitors which bind to or co-precipitate with nucleic acids during DNA extraction. In the present study we report a modified sodium dodecyl sulfate/phenol protocol that includes elimination of lysis solution to the bacteria cell wall and the washing of pellet with ethanol. The present study the relation A260/A280 and A260/A230 were of 1.84 ? 0.17 and 1.92 ? 0.62, respectively. These results showed that the DNA fraction is pure and may be used for PCR analysis future. To confirm this, the DNA?s purity was evaluated across a PCR amplification of fragment of the 16S gene using cell?s biomass from native phosphate solubilising bacteria (gram positive). Finally, the advantages of this procedure is that in the DNA extraction is not necessary the use of Proteinase K, and sonication to lyse samples prior to or in conjunction with lysis solutions.

Keywords : DNA extraction, Native phosphate solubilising bacteria, Gram positive, PCR.