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High-Throughput Cultivation of Bacterial Clones

red bacteria; however, mid-log phase growth and final cell density were equivalent to plate-cultured bacteria. No 550-nm absorption above background was observed in noninoculated wells of the plate.

A sample from every well of the plate was streaked onto an LB-ampicillin agar plate with a sterile toothpick and the plate was incubated at 37C for 24 hours. Bacterial growth was observed only from wells initially inoculated with bacteria. Bacterial growth was not observed in samples retrieved from noninoculated wells (data not shown). Therefore, cross contamination to alternate noninoculated wells did not occur in the aerated and nonaerated wells of the plate.

Plasmid Yield from Plate- and Flask-Cultured Bacteria

Plasmid DNA was isolated from 1 ml of plate- (aerated and nonaerated wells) and flask-cultured bacteria collected at the end of the incubation time period using the StrataPrep plasmid miniprep kit. Plasmid DNA yields were equivalent in plate- and flask-cultured bacteria, as judged by gel electrophoresis (Figure 3).

Fig.3

Conclusions

The Tempest thermoshaker allows 192 bacterial clones to be cultivated simultaneously in two, 96-deep-well plates. A special lid seals each well and prevents cross contamination. Bacterial growth rate, final cell density, and plasmid yield are virtually identical in aerated and nonaerated plate- and flask-cultured bacteria. The Tempest thermoshaker is a convenient and valuable tool when high-throughput capability is required.

REFERENCES

  1. Miller, J.H. (1972) Experiments in Molecular Genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York. pp. 32-33.


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