Difference between revisions of "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)
Line 7: Line 7:
 
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==mm/dd/yyyy==
 
==mm/dd/yyyy==
* Insert content here...
+
RhlI
 
+
{| {{table}}
 
+
|-
 +
|OD650||GFP in LB||pH
 +
|-
 +
|0.612||9929||7-8
 +
|-
 +
|}<br>
 +
EsaI
 +
{| {{table}}
 +
|-
 +
|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.<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.
 +
<br>
 +
'''11:40am'''
 +
<br>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
 +
{| {{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
 +
|-
 +
|}
 +
<br><br>
 +
minipreps picked 9/18/13
 +
{| {{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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|}
 
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__NOTOC__
 
__NOTOC__

Revision as of 14:10, 14 October 2013

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mm/dd/yyyy

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.