Team:Toronto/Notebook-w11-tue

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Tuesday, July 26

Wednesday, 7/27
Members Present: Celine, Kat, Karim, Bohdan, Hamed
LAB:
Morning:
Miniprepped overnight cultures that were obtained from single colonies that emerged from transformation following ligation.
Nanodropped the miniprep samples:
A
B
C
D
1
SAMPLE260/280260/230CONC (ng/uL)
2
RFP+ A11.910.849
3
RFP + A21.560.9111.1
4
RFP+A31.640.9214.3
5
RFP+ B12.240.8711.3
6
RFP+ B21.770.9513.1
7
RFP+ B31.780.9412.8
8
RFP-A11.651.0618
9
RFP- A21.790.8210.3
10
RFP-B1.680.879.9
11
Kit + B1.640.9915.7
12
CjBlue1.650.9416.8
Table1
Afternoon:
Ran gel on the miniprepped (undigested) samples
july26-laczligations.jpg
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Lane 1: RFP+ 1A Lane 2: RFP + A2 Lane 3: RFP+ A3 Lane 4: Ladder Lane 5: RFP+ B1 Lane 6: RFP + B2 Lane 7: RFP + B3 Lane 8: RFP - A1 Lane 9: Ladder Lane 10: RFP - A2 Lane 11: RFP-B Lane 12: Kit + B Lane 13: Cj blue
Dry run of the S30 sonication protocol. We tried two conditions:
3X10sec on/10sec off + 5 sec. (at amp 17)
4X 10 sec on/10sec off (amp 10)
RFP comparison.jpg
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Comparison of sonicated RFP (on the right) versus unsonicated RFP on the left. Note the colour change pre/post
RFP.jpg
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RFP after sonicating and centrifuging, brought up to the light.
RFP close up.jpg
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Close up of the sonicated and centrifuged RFP.
RFP CN tower.jpg
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Action shot of our RFP lysate and the CN tower.
TO DO:
For the next day:
Run gel on digested miniprepped samples (try digesting the kit plasmid with single and double enzymes)
PCR purify and 6hr DpnI digest of the gel purified backbone
Order more linearized backbone from iGEM