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

From OpenWetWare
Jump to: navigation, search
Owwnotebook icon.png Project name Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png

08/17/2015

AubR Testing AubR with 1μM 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 1μM synthetic N-Dodecanoyl-DL-homoserine lactone (C12-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)