Description
The mi-Bacterial Genomic DNA Kit is used to quickly and efficiently isolate high molecular weight genomic DNA (50-100 kb) from bacteria cells. This kit is based on the efficient release of genomic DNA by special cell lysis buffer rapid separation of genomic DNA from proteins, polysaccharides and lipids by selective adsorption of DNA to silica membrane. The DNA isolated can be used for all molecular biology applications (RFLP, PCR, Southern Blotting, etc). This method is fast, safe and simple for DNA preparation from different sources of samples.
Principle
The overall principle of DNA purification by silica-membrane technology is the specific and selective adsorption of DNA molecules to the silica-gel–membrane in the spin column. The cells are lysed during a short incubation with Proteinase K in the presence of chaotropic salt which immediately inactives all nucleases and enhances the binding of DNA. After the binding of the DNA in optimized high-salt buffers to the surface, all contaminants like primers, nucleotides, enzymes, mineral oil, salts, agarose, ethidium bromide, and other impurities from DNA samples, can be removed through specialized wash buffers. The purified DNA can be eluted by low-salt buffer or distilled water.
Time required
Less than 1 hour for gram - bacteria
Less than 2 hour for gram + bacteria
Advantages
Rapid isolation of genomic DNA from both, gram – (1h) and gram+ bacteria (2h)
Easy protocol
High efficiency
High yields:
Gram + bacteria : Staphylococcus epidermis (4 x 108 cells/ml) : 5-12ug
Gram - bacteria : Escherichia coli JM109 (2 x 109 cells/ml) : 20-25ug
High reproducibility
High molecular weight genomic DNA: 50-100 kb
The isolated DNA can be used for all molecular biology applications (RFLP, PCR, Southern Blotting, etc).
Final volume of DNA : 50 µl
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