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

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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 ligase1ul
Total38ul


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


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