Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2013/09/19

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(Autocreate 2013/09/19 Entry for Haynes_Lab:Notebook/Characterizing_AHL_quorum_sensing_homologs)
(mm/dd/yyyy)
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==mm/dd/yyyy==
==mm/dd/yyyy==
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* Insert content here...
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RhlI
-
 
+
{| {{table}}
-
 
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|-
 +
|OD650||GFP in LB||pH
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|-
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|0.612||9929||7-8
 +
|-
 +
|}<br>
 +
EsaI
 +
{| {{table}}
 +
|-
 +
|OD650||GFP in LB||pH
 +
|-
 +
|0.551||257||7-8
 +
|-
 +
|}
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Double checked super weak EsaI GFP expression and it's correct. Pelleted cells were not as green as RhlI.<br>
 +
F2620-RFP
 +
{| {{table}}
 +
|-
 +
|OD650||mCherry in LB
 +
|-
 +
|0.544||668
 +
|-
 +
|}<br>
 +
F2620-GFP
 +
{| {{table}}
 +
|-
 +
|OD650||GFP in LB
 +
|-
 +
|0.502||1166
 +
|-
 +
|}
 +
<br>
 +
Control supernatant was DH5αT K092900-E0420 which was not a perfect control. pH was 8-9.
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<br>
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'''11:40am'''
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<br>measured mCherry Ex:572/Em:607
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Spun Rhl and Esa at 10:30am. Moved 1mL to round bottom tubes. Added 40uL LuxR-GFP or LuxR-RFP. Added 1ul amp.
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RhlI
 +
{| {{table}}
 +
|-
 +
|Sender||Receiver
 +
|-
 +
|RhlI||LuxR-GFP
 +
|-
 +
|EsaI||LuxR-GFP
 +
|-
 +
|Ctrl||LuxR-GFP
 +
|-
 +
|RhlI||LuxR-RFP
 +
|-
 +
|EsaI||LuxR-RFP
 +
|-
 +
|Ctrl||LuxR-RFP
 +
|-
 +
|}
 +
<br>
 +
Thawed 1mL supernatant tubes, transferred to round bottom tubes. Added 40uL LuxR-GFP. Added 1uL amp.
 +
{| {{table}}
 +
|-
 +
|Sender||Receiver
 +
|-
 +
|RhlI 1||LuxR-GFP
 +
|-
 +
|RhlI 2||LuxR-GFP
 +
|-
 +
|RhlI 3||LuxR-GFP
 +
|-
 +
|}
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<br><br>
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minipreps picked 9/18/13
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{| {{table}}
 +
|-
 +
|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
 +
|-
 +
|}
 +
<br>
 +
pretty sure i put these in the freezer and did them another day.
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__NOTOC__
__NOTOC__

Revision as of 18:10, 14 October 2013

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mm/dd/yyyy

RhlI

OD650GFP in LBpH
0.61299297-8

EsaI

OD650GFP in LBpH
0.5512577-8

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

OD650mCherry in LB
0.544668

F2620-GFP

OD650GFP in LB
0.5021166


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

SenderReceiver
RhlILuxR-GFP
EsaILuxR-GFP
CtrlLuxR-GFP
RhlILuxR-RFP
EsaILuxR-RFP
CtrlLuxR-RFP


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

SenderReceiver
RhlI 1LuxR-GFP
RhlI 2LuxR-GFP
RhlI 3LuxR-GFP



minipreps picked 9/18/13

PartResult
K081002grew slowly
I13522-3grew slowly
K092900-E0420 (80.5)didn't grow
F2620-GFPgrew
RhlIgrew
F2620-RFPgrew
K081020grew
EsaIgrew
K092900-E0420 (80)grew


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


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