Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2013/05/29

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(Entry title)
Current revision (11:53, 30 May 2013) (view source)
(Entry title)
 
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Also picked TraI, TraR, and Ernesto's ligation of K092900-E0240 into BL21 for mini preps.
Also picked TraI, TraR, and Ernesto's ligation of K092900-E0240 into BL21 for mini preps.
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Came in at 8PM. BL21 senders (LuxI, LasI, RhlI) cultures weren't cloudy but TraI in DH5aT was cloudy. Could possibly be that BL21s grow slower and the volume is large (6ml). RhlR-GFP did not grow. LuxR-GFP grew. Sealed LuxR-GFP, TraI, and TraR sender ctrl and put them at 4deg.
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Picked RhlI, LuxI, and LasI into 2ml cultures to see if they would grow at smaller volumes. Also picked another RhlR-GFP just to see.
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Spun TraI and TraR and froze pellets to miniprep later. Ernesto's ligation of K092900-E0240 into BL21 did not grow. Left it shaking.
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Set up experiment to test plate reader with GFP receivers. Started shaking at 11am.

Senders (6ml) Receivers (2ml)
LuxILuxR-GFP
LasI
RhlI RhlR-GFP (maybe)
TraI
Controls
Positive (Kind of)GFP only (insert part #)
Negative sender TraR
Negative receiver none



Also picked TraI, TraR, and Ernesto's ligation of K092900-E0240 into BL21 for mini preps.



Came in at 8PM. BL21 senders (LuxI, LasI, RhlI) cultures weren't cloudy but TraI in DH5aT was cloudy. Could possibly be that BL21s grow slower and the volume is large (6ml). RhlR-GFP did not grow. LuxR-GFP grew. Sealed LuxR-GFP, TraI, and TraR sender ctrl and put them at 4deg.

Picked RhlI, LuxI, and LasI into 2ml cultures to see if they would grow at smaller volumes. Also picked another RhlR-GFP just to see.
Spun TraI and TraR and froze pellets to miniprep later. Ernesto's ligation of K092900-E0240 into BL21 did not grow. Left it shaking.


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