Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2013/10/23

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mm/dd/yyyy

miniprepped J06702 and the ligation colony I picked (backbone control plate had as many colonies as the ligation). Good yield for J06702, ok for other.

plasmid 260 280 260/280 concentration
J06702 0.247 0.128 1.93 247.2
ligation 0.071 0.035 2.006 70.9


Digested J06702 with

E only for 10 minutes
X only for 10 minutes
E and X for 10 minutes
E only for 5, then add X for 5 more minutes


Digested ligation with

P only
E/P


Ran on gel

Cut J06702 should be around 3kb. Possibly uncut, hence the crescent shapes and multiple bands.
pre-extraction



Extracted the bottom bands from the two J06702 cut with E/X
Arrows show where the bands were extracted.

post-extraction



Zymo gel recover kit

plasmid 260 280 260/280 concentration
J06702 3 0.017 0.011 1.468 16.729
J06702 4 0.008 0.005 1.583 7.782