Evaluation three methods of the extraction and purification of bacterial DNA of Gram positive and Gram negative bacteria

Abstract

Three methods were evaluated to choose the rapid and simple method of DNA extraction that is suitable for PCR detection. Ten isolates of Gram positive and Gram negative bacteria (Bacillus subtilis, Staphylococcus aureus, S. epidermidis, Streptococcus pyogenes, S. pneumonia: Escherichia coli, Klebsiella pneumoniae, Salmonella typhi, S. typhimurium and Proteus mirabilis). Gram positive bacteria were more resistant to cellular lyses due to high concentration of peptidoglycan within bacterial cell wall. The qualitative assessment of bacterial DNA revealed that DNA extraction with QuickExtract™ Bacterial DNA Extraction Kit method produced the highest DNA purity and highest DNA yield as compared with the other two methods. In conclusion, QuickExtract™ Bacterial DNA extraction kit method is technically simpler and more rapid than Wizard Genomic DNA purification kit and phenol- chloroform method which was more slow and low purity of yielded DNA. QuickExtract™ Bacterial DNA extraction kit is the best method for extracting genomic DNA from Gram positive and Gram negative bacteria, providing single-tube system. No toxic organic solvents, suitable for high-throughput applications, ready Lyse™ Lysozyme supplied and long DNA generated.