Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2015/08/18: Difference between revisions

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==08/17/2015==
==08/17/2015==
AubR
AubR
'''Testing AubR with 100μM synthetic N-buteryl-HHL (C4-HSL)'''
'''Testing AubR with XXXμM synthetic N-Dodecanoyl-DL-homoserine lactone (C12-HSL)'''


BjaR
BjaR
'''Testing BjaR with 100μM synthetic N-buteryl-HHL (C4-HSL)'''
'''Testing BjaR with 100nM synthetic N-buteryl-HSL (C4-HSL)'''<br>
BjaI produces isovaleryl-HSL but we can't buy it. According to the paper, it also responds to C4-HSL and C5-HSL at 100nM.
 


BraR
BraR
'''Testing BraR with 100μM synthetic N-buteryl-HHL (C4-HSL)'''
'''Testing BraR with 100nM synthetic N-(3-Oxotetradecanoyl)-L-homoserine lactone (3-oxo-C14-HSL or OC14-HSL)'''<br>


EsaR
EsaR
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LuxR
LuxR
'''Testing LuxR with 100μM synthetic N-buteryl-HHL (C4-HSL)'''
'''Testing LuxR with 1μM synthetic N-(3-oxohexanoyl)-HSL (aka N-(β-Ketocaproyl)-L-homoserine lactone aka 3O-C6-HSL)'''


RhlR
RhlR
'''Testing RhlR with 100μM synthetic N-buteryl-HHL (C4-HSL)'''
'''Testing RhlR with 100μM synthetic N-buteryl-HSL (C4-HSL)'''
:Diluted 1μL of 5mM C4-HSL into 99μL LB.
:Diluted 1μL of 5mM C4-HSL into 99μL LB.
:Put 2mL LB +amp into 14mL round bottom tube
:Put 2mL LB +amp into 14mL round bottom tube
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RpaR
RpaR
'''Testing RpaR with 100μM synthetic N-buteryl-HHL (C4-HSL)'''
'''Testing RpaR with XXXμM synthetic N-(p-Coumaroyl)-L-homoserine lactone (pC-HSL)'''
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Revision as of 22:38, 17 August 2015

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08/17/2015

AubR Testing AubR with XXXμM synthetic N-Dodecanoyl-DL-homoserine lactone (C12-HSL)

BjaR Testing BjaR with 100nM synthetic N-buteryl-HSL (C4-HSL)
BjaI produces isovaleryl-HSL but we can't buy it. According to the paper, it also responds to C4-HSL and C5-HSL at 100nM.


BraR Testing BraR with 100nM synthetic N-(3-Oxotetradecanoyl)-L-homoserine lactone (3-oxo-C14-HSL or OC14-HSL)

EsaR Testing EsaR with 1000μM synthetic N-(3-oxohexanoyl)-HSL (aka N-(β-Ketocaproyl)-L-homoserine lactone aka 3O-C6-HSL)

Diluted 1μL of 10mM C4-HSL into 99μL LB.
Put 2mL LB +amp into 14mL round bottom tube
Added 20μL diluted C4-HSL for a final concentration of 100μM C4-HSL
Added 100μL overnight culture of RhlR
Shook overnight

LuxR Testing LuxR with 1μM synthetic N-(3-oxohexanoyl)-HSL (aka N-(β-Ketocaproyl)-L-homoserine lactone aka 3O-C6-HSL)

RhlR Testing RhlR with 100μM synthetic N-buteryl-HSL (C4-HSL)

Diluted 1μL of 5mM C4-HSL into 99μL LB.
Put 2mL LB +amp into 14mL round bottom tube
Added 4μL diluted C4-HSL for a final concentration of 100μM C4-HSL
Added 100μL overnight culture of RhlR
Shook overnight

RpaR Testing RpaR with XXXμM synthetic N-(p-Coumaroyl)-L-homoserine lactone (pC-HSL)




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