Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2014/05/21

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(mm/dd/yyyy)
Current revision (19:56, 22 May 2014) (view source)
(mm/dd/yyyy)
 
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Ran above PCR on gel
Ran above PCR on gel
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Template'''
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| align="center" style="background:#f0f0f0;"|'''F primer'''
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| align="center" style="background:#f0f0f0;"|'''R primer'''
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| align="center" style="background:#f0f0f0;"|'''Expected band size'''
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| align="center" style="background:#f0f0f0;"|'''Actual band size'''
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|-
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| pTet-mCh||130||97||3130||3130
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|-
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| LuxI GFP rec||131||132||610||no band
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|-
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| LuxR 1:10 colony 1||103||114||1817||no band
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|-
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| LuxR 1:10 colony 2||103||114||1817||1817
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|-
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| LuxR 1:10 colony 3||103||114||1817||no band
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|-
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| LuxR 1:10 colony 4||103||114||1817||no band
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|-
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| LuxR 1:10 colony 5||103||114||1817||1817
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|-
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| LuxR 1:10 colony 5||103||107||1035||1035
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|}
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<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.  

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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
Bb13
RhlR23
EsaR14



Ran PCR, extension time 3:30 at 50°C

Template F primer R primer
pTet-mCh13097
LuxI GFP rec131132
LuxR 1:10 colony 1103104
LuxR 1:10 colony 2103104
LuxR 1:10 colony 3103104
LuxR 1:10 colony 4103104
LuxR 1:10 colony 5103104
LuxR 1:10 colony 5103107


Ran above PCR on gel

Template F primer R primer Expected band size Actual band size
pTet-mCh1309731303130
LuxI GFP rec131132610no band
LuxR 1:10 colony 11031141817no band
LuxR 1:10 colony 210311418171817
LuxR 1:10 colony 31031141817no band
LuxR 1:10 colony 41031141817no band
LuxR 1:10 colony 510311418171817
LuxR 1:10 colony 510310710351035



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



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