GEB's plasmid miniprep Kit is based upon a modified SDS-alkaline lysis of bacterial cells in combination with selective binding of the plasmid DNA to a special miniprep column. Each column has a binding capacity of at least 20 μg. This protocol provides a simple and reliable method to achieve the rapid isolation of highly purified DNA. The kit can also be used to isolate large recombinant constructs, such as BACs, PACs and P1s but some decrease in elution efficiency may be encountered.
The protocol has been optimized for bacterial cultures grown in LB (Luria-Bertani) broth, but can also be used for culture grown in rich broths. Each column can process up to 4 ml of bacterial culture grown in LB or up to 2 ml of culture grown in rich broth. The entire procedure can be completed in 20 min. The highly purified plasmid DNA is eluted in a small volume of this buffer eluent or deionized water and can be immediately used for many routine applications, such as DNA sequencing, restriction digestion, transcription in vitro, library screening, ligation and transformation, etc.