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Comparison of Growth Techniques and Media for the Purpose of Plasmid ,,, Isolation from E. coli Using the Eppendorf ,,, Perfectprep Plasmid Mini ,,, Kit

of Bacterial Cultures in 15 ml Conical Tubes and LidBac

A.

B.

C.

Figure 1. Comparison of E. coli cultivation in LB, 2X YT and TB mediums using 15 ml tubes and LidBac. A. Growth curves of bacterial cultures in LB media. B. Growth curves of bacterial cultures in 2X YT media. C. Growth curves of bacterial cultures in TB media.

In the experiments shown, the culture was allowed to proceed to the log phase. Figure 1 shows a comparison of cultures grown in 15 ml culture tubes and the Eppendorf LidBac in three different media. In all cases, growth curves of bacterial cultures were higher in the samples grown in LidBac. Clearly, the cultures experienced better growth when LidBac was used as the culture vessel, regardless of the media.

The table in Figure 2 demonstrates that the growth rates achieved were directly proportional to the yield of plasmid DNA after purification. A 1.4- to 3-fold increase is seen in these samples, depending on the media used. As expected, the greatest differences were seen in the samples grown in TB, which also had the greatest differences in growth curves (Figure 1C).

One of the critical factors related to the difference in growth rates is the superior air exchange characteristic of the hydrophobic 0.22 m membrane of LidBac. In contrast, the 15 ml culture tubes require loose capping to prevent contamination, thus limiting air exchange within the tube. Increased aeration and improved growth rates seen with the LidBac culture can also be attributed to the increased agitation rate that can be achieved with the Thermomixer R compared to conventional incubat
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