How to extract dna from gram positive bacteria


How to extract dna from gram positive bacteria

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AbstractGuanidine thiocyanate breakage of microorganisms has been the standard initial step in genomic DNA (gDNA) extraction of microbial DNA for two decades, despite the requirement how to extract dna from gram positive bacteria pretreatments to extract DNA from microorganisms other than Gram-negative bacteria. We report a quick and low-cost gDNA extraction protocol called EtNa that is efficient for bacteria and yeast over a broad range of concentrations. EtNa is based on a hot alkaline ethanol lysis. The solution can be immediately centrifuged to yield a crude gDNA extract suitable for PCR, or it can be directly applied to a silica column for purification.

Molecular biology techniques such as PCR, mass spectrometry, and sequencing have been optimized in order to speed up the detection and analysis of microorganisms ( 1). In this method, bacteria or yeasts are lysed directly by phenol and the supernatant is extracted with chloroform to remove traces of phenol. The supernatant contains DNA that is suitable for molecular analyses, such as PCR, restriction enzyme digestion and genomic library construction. This method is reproducible and simple for the routine DNA extraction from bacteria and yeasts.

DescriptionThe availability of simple DNA sample preparation methods is becoming increasingly important in fast-paced research settings, as the use of nucleic acid-based analyses continues to expand rapidly.




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