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Aurum Plasmid 96 Kit Tech Note, Rev A

agar plate and incubated overnight at 37C with shaking at 250 rpm. LB-ampicillin medium (500 ml) supplemented with 2% glycerol was then inoculated with 500 l of the starter culture and incubated overnight at 37C with shaking at 250 rpm. Culture density was measured by taking an absorbance reading of a 20-fold dilution of the culture at λ = 600 nm.

Plasmid Purification
Samples of a 4 OD culture were transferred to each well in columns 16 of six 96-well grow blocks. Similarly, samples of a 10 OD culture were transferred to each well in columns 712. The grow blocks were then centrifuged at 1,500 x g for 10 min, the medium decanted, and the grow blocks blotted on paper towels to remove residual medium. Plasmid pCMV was subsequently purified from the bacteria using a different manufacturers 96-well plasmid purification kit for each grow block and following each kits protocol. For all kits, the plasmid DNA was eluted or resuspended in 80 l of the appropriate elution buffer. Plasmid DNA concentrations and quality were measured using UV spectrophotometry. All samples from the 4 OD culture were run on agarose gels to confirm purity and concentration of the plasmid. For each manufacturers kit, 5 of the samples purified from the 4 OD culture were sequenced on an Applera ABI PRISM 3100 capillary array sequencer, and the resulting electropherograms were analyzed for ambiguous counts within the first 600 bases to assess relative comparative read lengths.


Results
Plasmid Yields and Concentrations
With respect to overall plasmid DNA mass yield, the Aurum plasmid 96 kit was one of the 2 best performers, giving the highest yield of all kits with the 4 OD culture
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