Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2015/07/01

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07/01/2015

Testing RhlR with 100μM synthetic N-buteryl-HHL (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



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



PCR pEsaR and LasR.
Ran gel for colony PCR of RhlI colonies. None of the colonies worked; next step is to pick more (since there were a lot on the plate) and try again before abandoning the batch.


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