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Qualitative and Quantitative Assessment of Plasmid DNA Isolation, Rev A

for 2 hr at 37C. Digests were resolved on 0.7% agarose TAE gels. Bacteria were transformed as described above. Yeast were transformed by either a standard lithium acetate protocol (Gietz and Woods 1994) or by electroporation with 100 ng of plasmid. DNA sequencing was performed directly on 100 ng of plasmid with 1 pmol of primer by a centralized core facility under standard conditions (available on request).

*Bacterial Growth Guidelines
Aurum plasmid mini and midi kits can process up to 12 ODml and 50 ODml, respectively, of bacterial culture grown in a variety of different broths, such as LB (Luria-Bertani broth), LBG (LB + 2% glycerol), TB (Terrific Broth), and 2x YT. The quantity of bacteria processed in a purification procedure, measured in ODml, was determined as follows:

First, a sample of culture was diluted 1:20 by combining 50 l of bacterial culture with 950 l of sterile broth. The optical density of this sample at λ = 600 nm (OD600) was multiplied by 20 to calculate the density of the original undiluted culture.

Depending upon the OD600, a specific volume of the undiluted culture was selected to provide an appropriate quantity of bacterial cells for processing. Appropriate volumes of culture were calculated using the following equation:

(OD600 of undiluted culture) x (culture volume in ml) = #ODml

For example, 12 ODml of bacteria would require 2 ml of an undiluted culture with an OD600 of 6, or 4 ml of a culture with an OD600 of 3.

1 OD600 is equivalent to appr
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