Description
mi-PCR Purification Kit is designed for rapid purification of PCR products combined with a extraordinarily high recovery rate.
DNA Binding Buffer is added to the PCR reaction and the mixture is applied to a spin column containing silica-based membranes where the double stranded DNA is selectively absorbed.
DNA polymerases, buffer, remaining primers, and dNTP are removed with alcohol-containing Column Wash Buffer. Since the DNA is eluted with nuclease-free distilled water or TE buffer, no precipitation is necessary. All solutions are passed through the spin column with brief centrifugations, allowing the completion of the protocol in < 10 minutes. This kit eliminates the use of hazardous chemical such as phenol and chloroform.
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.
Advantages
Rapid purification of PCR product (less than 10 min)
DNA size range : 100 bp – 15 kb
Recovery rates : up to 95%
Easy protocol
Final volume : 50 µl
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