( Cultur...)High-Throughput Cultivation of Bacterial Clones

Culture 192 clones simultaneously with the tempest thermoshaker

Jeff Braman
Stratagene

Figure 1: The tempest Thermoshaker

The tempest thermoshaker and a 96-deep-well round-bottom plate were used to cultivate bacterial clones in a high-throughput format. Early- and mid-log phase rates of bacterial growth were equivalent in aerated and nonaerated wells of the plate. Late-log phase growth rates of bacteria cultured in nonaerated wells were only slightly slower than growth rates measured in aerated wells. Final cell densities in aerated and nonaerated wells were identical. Flask-cultured bacteria demonstrated a slower initial rate of growth compared to plate-cultured bacteria, however, mid-log phase growth and final cell density were equivalent to plate-cultured bacteria. Well-to-well plate contamination was undetectable. Equal quantities of plasmid DNA were recovered from equal volumes of plate- and flask-cultured bacteria sampled at the end of the incubation period.

The Tempest thermoshaker is a compact benchtop incubator and shaker that is used for regulated temperature incubation (0 to 40C) and rotary mixing (200 to 1500 rpm) of multiple samples. The unit is supplied with a holder to accommodate fifty six 1.5-ml tubes. With Stratagenes minitube adapters, the thermoshaker can also hold 0.5- or 0.6-ml tubes. A microplate platform, also available, holds four, 96-well microtiter plates or two, 96-deep-well plates. The latter plates are particularly useful for cultivating bacteria in a high-throughput.

96-Well Plate vs. Flask Cultivation of Bacteria

Growth rates and final cell densities were measured for plate- and flask-cultured Epicurian Coli
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