Volume 15, Issue 45 (9-2014)
Zanko J Med Sci 2014, 15(45): 9-16 |
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Shahbazi B, Narenji H. Comparison of four methods of DNA extraction from gram-negative and gram-positive bacteria. Zanko J Med Sci 2014; 15 (45) :9-16
URL: http://zanko.muk.ac.ir/article-1-32-en.html
URL: http://zanko.muk.ac.ir/article-1-32-en.html
Abstract: (18594 Views)
Abstract
Background and Aim: DNA extraction is the first step in the genetic engineering research and obtains a suitable protocol for the preparation of a purified DNA which is essential. In this context, many manual and kit methods were provided. The aim of this study is to achieve a faster and low cost method for DNA extraction of Gram-positive bacteria and gram-negative bacteria. We compared four DNA extraction methods, Kit extraction, phenol chloroform, using detergent laundry brand tag and boiling.
Materials and Methods: In this study, bacterial suspensions of Staphylococcus aureus and Vibrio cholerae were produced similar to McFarland 0.5 turbidity. Bacterial DNA was extracted using four different methods and three times for each. Using spectrophotometer and gel electrophoresis were measured concentrations and quality of DNA extraction product accordingly. In order to evaluate the efficiency of DNA extraction method, PCR, ERIC PCR and REP-PCR were performed on some of housekeeping genes downstream regions.
Results: The concentration of the extracted DNA and results of PCR, ERIC-PCR and REP-PCR were different between gram-positive and gram-negative bacteria significantly (P< 0.05). ERIC PCR and REP-PCR efficiency of the boiling and chloroform extraction methods was disrupted only in ERIC PCR. No deficiency was observed in Sinagen kit method.
Conclusions: The findings of this study show that despite the availability and lower cost of manual methods, these methods can be substitute for kit method.
Type of Study: Research |
Subject:
Special
Received: 2014/07/19 | Accepted: 2014/09/3 | ePublished: 2014/09/16
Received: 2014/07/19 | Accepted: 2014/09/3 | ePublished: 2014/09/16
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