US EPA

2899591 

Journal Article 

Novel Approaches of Genomic DNA Isolation for Identification of Cultivable Bacteria 

Pindi, PK; Srinath, RR; Shanker, AS 

2013 

1 

Jundishapur Journal of Microbiology
ISSN: 2008-3645
EISSN: 2008-4161 

6 

10 

10.5812/jjm.8339 

WOS:000330519300018 

Background: Identification of novel pathogenic bacteria always has a crucial role in clinical microbiology to defeat uncontrolled diseases.



Objectives: Sample preparation platform has been developed for rapid and simple detection of infectious organisms from the point of care diagnostics and molecular manipulation by using three novel genomic DNA isolation protocols.



Materials and Methods: Most of the genomic DNA isolation protocols are represented with SDS, alkaline lysis with extraction buffer and phenol, organic chloroform extractions and then enzymatic purifications, but in our reconnoiter, we only used three single chemicals including ethanol, phenol and hydrogen peroxide which are major disinfectants and sterilizers, acting on the cell wall and cell membrane proteins, followed by denaturation of the bacterial cell and simultaneous lysis.



Results: This study used the mentioned three major chemicals in three different protocols to avoid unnecessary purification steps especially for gene amplification and diagnostics reports. Genomic DNA was recovered by adding 50 mu L double distilled water.



Conceivably, we applied this principle for three novel DNA extraction methods and succeeded to achieve purified high-yield DNA. Conclusions: Purified high-yield DNA was obtained for molecular manipulations and elimination of diagnostic concerns. 

Acinetobacter sp; Bacterial Culture; Ethanol; Hydrogen Peroxide; Phenol; Bacillus sp