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

From OpenWetWare
Jump to navigationJump to search
Line 24: Line 24:


<font color = purple>  
<font color = purple>  
PCR pEsaR and LasR  
PCR pEsaR and LasR. <br>
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.




Revision as of 17:06, 2 July 2015

Project name <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

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.



Retrieved from "https://openwetware.org/mediawiki/index.php?title=Haynes_Lab:Notebook/Characterizing_AHL_quorum_sensing_homologs/2015/07/01&oldid=891346"