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Tuesday, July 26

Wednesday, 7/27

Members Present: Celine, Kat, Karim, Bohdan, Hamed

LAB:

Morning:

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Miniprepped overnight cultures that were obtained from single colonies that emerged from transformation following ligation.

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Nanodropped the miniprep samples:

	A	B	C	D
1	SAMPLE	260/280	260/230	CONC (ng/uL)
2	RFP+ A1	1.91	0.84	9
3	RFP + A2	1.56	0.91	11.1
4	RFP+A3	1.64	0.92	14.3
5	RFP+ B1	2.24	0.87	11.3
6	RFP+ B2	1.77	0.95	13.1
7	RFP+ B3	1.78	0.94	12.8
8	RFP-A1	1.65	1.06	18
9	RFP- A2	1.79	0.82	10.3
10	RFP-B	1.68	0.87	9.9
11	Kit + B	1.64	0.99	15.7
12	CjBlue	1.65	0.94	16.8

Table1

Afternoon:

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Ran gel on the miniprepped (undigested) samples

july26-laczligations.jpg

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

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Dry run of the S30 sonication protocol. We tried two conditions:

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3X10sec on/10sec off + 5 sec. (at amp 17)

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4X 10 sec on/10sec off (amp 10)

RFP comparison.jpg

Comparison of sonicated RFP (on the right) versus unsonicated RFP on the left. Note the colour change pre/post

RFP.jpg

RFP after sonicating and centrifuging, brought up to the light.

RFP close up.jpg

Close up of the sonicated and centrifuged RFP.

RFP CN tower.jpg

Action shot of our RFP lysate and the CN tower.

TO DO:

For the next day:

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Run gel on digested miniprepped samples (try digesting the kit plasmid with single and double enzymes)

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PCR purify and 6hr DpnI digest of the gel purified backbone

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