Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2013/05/20: Difference between revisions

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Also transformed TraR and picked J06702 (mCherry)
Also transformed TraR and picked J06702 (mCherry) and more F2620


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Revision as of 12:06, 28 May 2013

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Entry title

Miniprep liquid culture of F2620 (received agar stabs from the registry).


Part number 260 280 260/280 ng/ul
F2620 miniprep 0.082 0.047 1.738 81.545


Ernesto cut some E0240 that I can use. Cut F2620 with E/S to insert in front of E0240.
Gel stopped running after what looked like about 5-10 minutes. I came back after an hour to find it flashing E1. I restarted the gel and it ran fine for the next hour.
The gel showed a strong band (with some extra bands) but the gel extraction yield was negative:

Part number 260 280 260/280 ng/ul
F2620 gel ext -0.002 0 -15 -1.578


Continued with the ligation. E0240 backbone concentration (cut with E/X) was 6ng/ul. Should need 10ul for 50ng but I used 5ul and 25ng.

Insert DNA 13ul
Backbone DNA 5ul
Roche 2X buffer (Behzad) 19ul
T4 DNA ligase 1ul
Total 38ul


Also transformed TraR and picked J06702 (mCherry) and more F2620


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