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Friday June 24, 2016

Friday, 6/24

Members Present: Hamed, Kat, Alex, Cathy, Dk, Tam

LAB:

Morning:

- Counting number of colonies in each plate for the second ccc (first batch of rubidium chloride chmically competent cells) transformation experiment (along with cca, ccb and negative controls). Note that CCA and CCB refer to the first two batches of competent cells produced with MgCl. CCC refers to the RbCl cells.

- ccc transformation was effective (better than cca and ccb); transformation efficiency to be calculated

0.5pg_23_06.jpg

Bacterial transformation with 0.5pg plated. 1 CFU on CCA plate.

	A	B	C	D
1	CC batch	ng of DNA plated	CFU	Transformation efficiency
2	CCA	9.92 x 10^-6	1	1.01 x 10^8 CFU/ug
3	CCB	9.92 x 10^-6	0	0
4	CCC	9.92 x 10^-6	0	0

Table1

10pg_23_06.jpg

Bacterial transformation with 10pg plated. 4 CFU with CCA, 5 CFU with CCB, and 15 CFU with CCC.

	A	B	C	D
1	CC batch	ng of DNA plated	CFU	Transformation efficiency
2	CCA	1.98 x 10^-3 ng	4	2.02 x 10^6 CFU/ug
3	CCB	1.98 x 10^-3 ng	5	2.52 x 10^6 CFU/ug
4	CCC	1.98 x 10^-3 ng	15	1.57 x 10^6 CFU/ug

Table2

50pg_23_06.jpg

Bacterial transformation with 50pg of DNA plated. 18 CFU on CCA, 11 CFU on CCB, 51 CFU on CCC.

	A	B	C	D
1	CC batch	ng of DNA plated	CFU	Transformation efficiency (CFU/ug)
2	CCA	9.92 x 10^-3	18	1.81 x 10^6
3	CCB	9.92 x 10^-3	11	1.11 x 10^6
4	CCC	9.92 x 10^-3	50	5.04 x 10^6

Table3

RFP1A_23_06.jpg

Bacterial transformation with RFP1A at a concentration of 16.7ng/uL. 574 CFU on CCA, 800 CFU on CCB, 1600 CFU on CCC.

	A	B	C	D
1	CC batch	ng of DNA plated	CFU	Transformation efficiency (CFU/ug)
2	CCA	3.31	574	1.73 x 10^5
3	CCB	3.31	800	2.42 x 10^5
4	CCC	3.31	1600	4.83 x 10^5

Table4

no plasmid 23_06.jpg

Control with no plasmid added. No CFU on CCA, CCB, CCC.

LB control_23_06.jpg

Control - grew CCC on LB without CAM to ensure that flash frozen cells were still viable (they were)

Afternoon:

- PCR amplification of the backbone and _Short Linear TetO-LacZ-alpha and measuring DNA concentration for each using nanodrop:

nanodrop backbone.jpg

Nanodrop of the PCR amplified pSB1C3 backbone from RFP

Nanodrop of the PCR amplified Short Linear Tet0-LacZ alpha

nanodrop of short linear TetO-LacZ alpha.jpg

TO DO:

For the next day:

LAB TEAM:

LAB MANAGERS:

Gmail correspondence:

Team:Toronto/Notebook-w06-fri