Difference between revisions of "Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2014/05/21"

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(mm/dd/yyyy)
(mm/dd/yyyy)
Line 44: Line 44:
 
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<br>
 
Ran above PCR on gel
 
Ran above PCR on gel
 
+
{| {{table}}
 +
| align="center" style="background:#f0f0f0;"|'''Template'''
 +
| align="center" style="background:#f0f0f0;"|'''F primer'''
 +
| align="center" style="background:#f0f0f0;"|'''R primer'''
 +
| align="center" style="background:#f0f0f0;"|'''Expected band size'''
 +
| align="center" style="background:#f0f0f0;"|'''Actual band size'''
 +
|-
 +
| pTet-mCh||130||97||3130||3130
 +
|-
 +
| LuxI GFP rec||131||132||610||no band
 +
|-
 +
| LuxR 1:10 colony 1||103||114||1817||no band
 +
|-
 +
| LuxR 1:10 colony 2||103||114||1817||1817
 +
|-
 +
| LuxR 1:10 colony 3||103||114||1817||no band
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|-
 +
| LuxR 1:10 colony 4||103||114||1817||no band
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|-
 +
| LuxR 1:10 colony 5||103||114||1817||1817
 +
|-
 +
| LuxR 1:10 colony 5||103||107||1035||1035
 +
|}
 
<br><br>
 
<br><br>
 
picked LuxI and RhlI GFP senders into 4mLs media and shook overnight to screen for successful receiver transformations.  
 
picked LuxI and RhlI GFP senders into 4mLs media and shook overnight to screen for successful receiver transformations.  

Revision as of 15:56, 22 May 2014

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mm/dd/yyyy

Golden gate 1:10 RhlR and EsaR receivers grew but there were colonies on the backbone plate

Ligation Colonies
Bb 13
RhlR 23
EsaR 14



Ran PCR, extension time 3:30 at 50°C

Template F primer R primer
pTet-mCh 130 97
LuxI GFP rec 131 132
LuxR 1:10 colony 1 103 104
LuxR 1:10 colony 2 103 104
LuxR 1:10 colony 3 103 104
LuxR 1:10 colony 4 103 104
LuxR 1:10 colony 5 103 104
LuxR 1:10 colony 5 103 107


Ran above PCR on gel

Template F primer R primer Expected band size Actual band size
pTet-mCh 130 97 3130 3130
LuxI GFP rec 131 132 610 no band
LuxR 1:10 colony 1 103 114 1817 no band
LuxR 1:10 colony 2 103 114 1817 1817
LuxR 1:10 colony 3 103 114 1817 no band
LuxR 1:10 colony 4 103 114 1817 no band
LuxR 1:10 colony 5 103 114 1817 1817
LuxR 1:10 colony 5 103 107 1035 1035



picked LuxI and RhlI GFP senders into 4mLs media and shook overnight to screen for successful receiver transformations.