Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2013/07/19

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(07/19/2013)
Current revision (23:49, 21 July 2013) (view source)
(07/19/2013)
 
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Picked one colony from each of the positive ones and K091134 because I don't have another LasR option.<br>
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Transformed each of the positive ones and K091134 because I don't have another LasR option.<br>
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<br>Transformed the second annealing reaction.

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07/19/2013

Ligation/transformation plates have colonies but they're all in the middle. Same with the backbone control. Picked colonies just in case.
I03522 or whatever has colonies. Picked two.
Started the oligo annealing over again. I shouldn't have cut or dephosphorylated the oligos. That was dumb.
Miniprepped the colonies I picked last night. Tried splitting the 3ml cultures into 2-1.5ml cultures to mini prep. Still got low yields.

PartSizeMiniprep yield 1Miniprep yield 2
K092900+cyan~2900
F2620+GFP~2000
K082035 68
K084007835


Cut a bunch of QS plasmids from the last few months with X/P to see which ones might be worth transforming and growing up to PCR out parts for the expression vectors. Ran on a gel

No. NameLengthGel check
1LuxR-GFP Receiver 1900 +
2K092900 + E02402200-
3 C0061 - 1643couldn't tell
4C0061 - 2643couldn't tell
5C0062781couldn't tell
6F2620 (81ng/ul)1061+
7F2620 (52ng/ul)1061+
8K0911342048-
9K0929002102_
10K145254862+
11K082035884+
12I0466942+
13K084007835+
14RBS-LasI600+
15messed up?-
16K092900-cyan2900-
17F2620-GFP2000-
18K084007835+
19RBS-LuxI798not sure
20J06702 56ng/ul869+


Transformed each of the positive ones and K091134 because I don't have another LasR option.

Transformed the second annealing reaction.



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