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Simple Purification of DNA from Plasmid Minipreps, PCR Amplifications,,,and Agarose Gels

ash and spin
  • Add elution buffer and spin to collect DNA
  • Figure 1 outlines the simple and rapid protocols used for purifying miniprep plasmid DNA, PCR products and agarose gel-fractionated DNA. Each microspin cup in the StrataPrep plasmid miniprep kit yields up to 20 g of purified plasmid DNA from overnight bacterial cultures of 1.5 to 3 ml. Each microspin cup in the StrataPrep PCR purification kit and DNA gel extraction kit yields as much as 10 g of purified DNA.

    Plasmid Purification

    figure 2

    Plasmid DNA isolated with the StrataPrep plasmid miniprep kit exhibits OD260 -to-OD280 ratios between 1.8 and 2.0, indicating a high degree of nucleic acid purity.2 Starting with 30 individual 1.5- to 3-ml liquid cultures, purified plasmid DNA can be obtained in an average of 2.5 minutes per miniprep. Plasmid and cosmid DNAs isolated with the StrataPrep plasmid miniprep kit are easily digested with restriction enzymes (figure 2). Plasmid DNA purified with the StrataPrep kit transforms chemically competent E. coli with efficiencies equivalent to those obtained for DNA that has been purified using cesium chloride density gradients. For example, 100 pg of a 2.9-kb plasmid purified using either the StrataPrep kit or the cesium chloride density gradient method transformed Epicurian Coli XL2-Blue ultracompetent cells with an efficiency of 2 x 109 colony forming units (cfu)/g of plasmid DNA (data not shown).


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