Difference between revisions of "Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2013/09/19"

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==09/19/2013==
 
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Revision as of 16:16, 2 December 2013

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09/19/2013

RhlI

OD650 GFP in LB pH
0.612 9929 7-8

EsaI

OD650 GFP in LB pH
0.551 257 7-8

Double checked super weak EsaI GFP expression and it's correct. Pelleted cells were not as green as RhlI.
F2620-RFP

OD650 mCherry in LB
0.544 668

F2620-GFP

OD650 GFP in LB
0.502 1166


Control supernatant was DH5αT K092900-E0420 which was not a perfect control. pH was 8-9.
11:40am
measured mCherry Ex:572/Em:607 Spun Rhl and Esa at 10:30am. Moved 1mL to round bottom tubes. Added 40uL LuxR-GFP or LuxR-RFP. Added 1ul amp. RhlI

Sender Receiver
RhlI LuxR-GFP
EsaI LuxR-GFP
Ctrl LuxR-GFP
RhlI LuxR-RFP
EsaI LuxR-RFP
Ctrl LuxR-RFP


Thawed 1mL supernatant tubes, transferred to round bottom tubes. Added 40uL LuxR-GFP. Added 1uL amp.

Sender Receiver
RhlI 1 LuxR-GFP
RhlI 2 LuxR-GFP
RhlI 3 LuxR-GFP



minipreps picked 9/18/13

Part Result
K081002 grew slowly
I13522-3 grew slowly
K092900-E0420 (80.5) didn't grow
F2620-GFP grew
RhlI grew
F2620-RFP grew
K081020 grew
EsaI grew
K092900-E0420 (80) grew


pretty sure i put these in the freezer and did them another day.