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Protocol Name
DOI:10.17504/protocols.io.gwnbxde RRID Copied  
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Lawrence A. Klobutcher 2017. Increased sensitivity of Euplotes crassus to selective agents using 0.3 M glucose-based culture conditions.. protocols.io dx.doi.org/10.17504/protocols.io.gwnbxde
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Protocol Information

URL: https://dx.doi.org/10.17504/protocols.io.gwnbxde

Authors: Lawrence A. Klobutcher

Group: Protist Research to Optimize Tools in Genetics (PROT-G)

Summary: Numerous unpublished studies indicate that Euplotes crassus is resistant to extremely high concentrations of commonly used selective agents when grow in artificial seawater. We have found that when E. crassus is grown in 1 part artificial seawater + 9 parts 0.3 M glucose, they are much more sensitive to a number of selective agents. For example, 1080 ug/ml paromomycin in artificial seawater only inhibited growth, as little as 120 ug/ml was effective in killing cells when grown with 0.3 M glucose. Similarly, G418 at 400 ug/ml had little effect in seawater, but as little as 100 ug/ml blocked growth in 0.3 M glucose. This may not prove true for all selective agents, as we saw no effect with paclitaxel up to 100 uM under either growth condition.

Affiliations: University of Connecticut Health Center

Version: 1

Publication Date: 2017

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Source: Protocols.io