Hızlı Bakteriyel DNA Ekstraksiyon Metotlarının Karşılaştırılması

Bu çalışma bakteriyel DNA ekstraksiyonunda kullanılan hızlı metotları karşılaştırmak amacıyla yapıldı. Bu amaçla Gram pozitif bakterilere örnek olarak Staphylococcus sp., Gram negatiflere örnek olarak Escherichia coli seçildi. Bu bakterilerin buyyon kültürlerinden (~107 cfu/ml) kaynatma, sonikasyon ve dondurma-çözme yöntemleri ile DNA ekstraksiyonu yapıldı. DNA miktarları 260 nm dalga boyunda spektrofotometre ile okundu. Sonikasyon ile elde edilen DNA miktarı diğer yöntemlerle elde edilen DNA miktarlarından istatistiksel olarak daha fazla bulundu (p

Anahtar Kelimeler:

Staphylococcus sp, Escherichia coli , DNA ekstraksiyonu

Comparison Of Rapid Bacterial DNA Extraction Methods

This study was carried out to compare rapid bacterial DNA extraction methods. Strains of Staphylococcus sp. and E. coli were preferred to represent Gram positive and Gram negative bacteria, respectively. DNA extractions from the cultured bacteria (~ 108 cfu/ml) were made by the methods of boiling, sonication and freze-thawing. Then, the quantities of DNA extracted were measured by spectrophotometric analysis at 260nm. DNA quantities obtained by sonication method were statistically more than those obtained by other methods (p<0.05)

Keywords:

Staphylococcus sp, Escherichia coli , DNA extraction,

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Maslow, J. N., Mulligan, M. E., Arbeit, R. D. Molecular Epidemiology: Application of Contemporary Techniques to the Typing of Microorganisms. Clin. Infect Dis. 17: 64-153 (1993).
van Belkum, A. DNA Fingerprinting of Medically Important Microorganisms by Use of PCR. Clin. Microbiol. Rev. 7: 84-174 (1995).
Wang, G., Whittam, T. S., Berg, C. M., Berg, D.E. RAPD (Arbitrary Primer) PCR is More Sensitive Than Multilocus Enzyme Electrophoresis for Distinguishing Related Bacterial Strains. Nucleic Acids Res. 21: 5930-5933 (1993).
Kevin, A., Comparison of Rapid Methods for the Extraction of Bacterial DNA from Colonic and Fecal Lumen Contents of the Pig. J. Applied Microbiol. 94: 988-993 (2003).
Rengarajen, K., Cristol, M.S., Mehta, M., Nickerson, J.M. Quantifying DNA Concentrations Using Fluorometry: A Comparison of Fluorophores. Molecular Vision. 8: 416-421 (2002).
Maniatis, T., Sambrook, J. Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory Pres, USA. (1989).
Newton, C.R. PCR Essential Data. John Wiley and Sons LTD. Baffins Lane, Chicheter, West Sussex PO 19, 1UD, UK. (1995).
More, M.I., Herrick, J.B., Silva, M.C., Ghiorse, W.C., Madsen, E.L. Quantitative Cell Lysis of Indigenous Microorganism and Rapid Extraction of Microbial DNA from Sediment. Applied and Environmental Microbiology 60:5, 1572-1580 (1994).
Yeates, C., Gillings M. R., Davison, A. D., Altavilla, N., Veal, D. A. Methods for Microbial DNA Extraction from Soil for PCR Amplification. Biological Procedures Online. 1:40-47 (1998).
0. Pitcher, D. G., Saunders, N. A., Owen R. J. Rapid Extraction of Bacterial Genomic DNA with Guanidium Thiocyanate. Letters in Applied Microbiology 8:151-156 (1989).