Team:ZJU-China/Project/ElvesCooperation/Results

From 2013.igem.org

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Result:
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Characterize Chemotaxis of  Detectors
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Characterize Communication Between Detectors And Cleaners
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Characterize the Function of Cleaners
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Characterize Chemotaxis of Detectors
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The strain E.coli JW1870 (ΔcheZ) ,which was a generous gift from Prof. John S. Parkinson, University of Utah ,was mainly used for our experiments. The bacteria containing Plasmid 1 was incubated in LB medium with 100ug/mL Amp until mid-log phage. 10 uL suspension culture was diluted with 1mL LB medium and then dripped at the center of a semi-solid plate, which contained 0.2% agar , 100ug/mL Amp and was covered with 2mM atrazine on the surface. The plate was incubated at 37℃ for 10hrs. The negative control was required.
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Characterize Communication Between Detectors And cleaners
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As what has been illustrated in Design part, Detector contains plasmid 1 and plasmid 2 while Cleaner contains plasmid 3.
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Since GFP expression in Detector has been proved and detected in Characterization of Atrazine riboswitch. We directly verify the communication between Detectors and Cleaners by observing GFP and RFP with confocal microscope.
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1、LB media cultivation
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Both Detector and Cleaner were incubated in LB media to mid-log phage (OD600 reached 0.6 approximately) and then was induced by 0.2mM IPTG and 2mM atrazine simultaneously. Further incubation was conducted at 37℃ /180 rpm for 8hrs.
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The following images were photographed with confocal microscope after incubation. Negative control without Atrazine was still conducted.

Revision as of 01:55, 28 September 2013

Elves' Cooperation: Result

Result: Characterize Chemotaxis of Detectors Characterize Communication Between Detectors And Cleaners Characterize the Function of Cleaners

Characterize Chemotaxis of Detectors The strain E.coli JW1870 (ΔcheZ) ,which was a generous gift from Prof. John S. Parkinson, University of Utah ,was mainly used for our experiments. The bacteria containing Plasmid 1 was incubated in LB medium with 100ug/mL Amp until mid-log phage. 10 uL suspension culture was diluted with 1mL LB medium and then dripped at the center of a semi-solid plate, which contained 0.2% agar , 100ug/mL Amp and was covered with 2mM atrazine on the surface. The plate was incubated at 37℃ for 10hrs. The negative control was required.

Characterize Communication Between Detectors And cleaners As what has been illustrated in Design part, Detector contains plasmid 1 and plasmid 2 while Cleaner contains plasmid 3.

Since GFP expression in Detector has been proved and detected in Characterization of Atrazine riboswitch. We directly verify the communication between Detectors and Cleaners by observing GFP and RFP with confocal microscope.

1、LB media cultivation Both Detector and Cleaner were incubated in LB media to mid-log phage (OD600 reached 0.6 approximately) and then was induced by 0.2mM IPTG and 2mM atrazine simultaneously. Further incubation was conducted at 37℃ /180 rpm for 8hrs. The following images were photographed with confocal microscope after incubation. Negative control without Atrazine was still conducted.