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

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==mm/dd/yyyy==
==07/19/2013==
* Insert content here...
Ligation/transformation plates have colonies but they're all in the middle. Same with the backbone control. Picked colonies just in case. <br>
I03522 or whatever has colonies. Picked two. <br>
Started the oligo annealing over again. I shouldn't have cut or dephosphorylated the oligos. That was dumb. <br>
Miniprepped the colonies I picked last night. Tried splitting the 3ml cultures into 2-1.5ml cultures to mini prep. Still got low yields.  
{| {{table}}
|-
|Part||Size||Miniprep yield 1||Miniprep yield 2
|-
|K092900+cyan||~2900||
|-
|F2620+GFP||~2000||
|-
|K082035|| || 6||8
|-
|K084007||835||
|-
|}
<br>
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<br>
 
{| {{table}}
|-
|No. || Name||Length||Gel check
|-
|1||LuxR-GFP Receiver|| 1900|| +
|-
|2||K092900 + E0240||2200||-
|-
|3 ||C0061 - 1||643||couldn't tell
|-
|4||C0061 - 2||643||couldn't tell
|-
|5||C0062||781||couldn't tell
|-
|6||F2620 (81ng/ul)||1061||+
|-
|7||F2620 (52ng/ul)||1061||+
|-
|8||K091134||2048||-
|-
|9||K092900||2102||_
|-
|10||K145254||862||+
|-
|11||K082035||884||+
|-
|12||I0466||942||+
|-
|13||K084007||835||+
|-
|14||RBS-LasI||600||+
|-
|15||messed up||?||-
|-
|16||K092900-cyan||2900||-
|-
|17||F2620-GFP||2000||-
|-
|18||K084007||835||+
|-
|19||RBS-LuxI||798||not sure
|-
|20||J06702 56ng/ul||869||+
|-
|}
<br>
Transformed each of the positive ones and K091134 because I don't have another LasR option.<br>
<br>Transformed the second annealing reaction.  




Revision as of 20:49, 21 July 2013

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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.

Part Size Miniprep yield 1 Miniprep yield 2
K092900+cyan ~2900
F2620+GFP ~2000
K082035 6 8
K084007 835


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. Name Length Gel check
1 LuxR-GFP Receiver 1900 +
2 K092900 + E0240 2200 -
3 C0061 - 1 643 couldn't tell
4 C0061 - 2 643 couldn't tell
5 C0062 781 couldn't tell
6 F2620 (81ng/ul) 1061 +
7 F2620 (52ng/ul) 1061 +
8 K091134 2048 -
9 K092900 2102 _
10 K145254 862 +
11 K082035 884 +
12 I0466 942 +
13 K084007 835 +
14 RBS-LasI 600 +
15 messed up ? -
16 K092900-cyan 2900 -
17 F2620-GFP 2000 -
18 K084007 835 +
19 RBS-LuxI 798 not sure
20 J06702 56ng/ul 869 +


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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