Revision as of 10:38, 15 September 2015

Project Results

Competent Cell Test Kit, RFP construct (iGEM)

Before using our competent E. coli TOP10 cells in the important experiments, we used the Competent Cell Test Kit to test the efficiency of our competent cells!

The kit includes five vials of each different DNA concentration: 50pg/μl, 20pg/μl, 10pg/μl, 5pg/μl, 0.5pg/μl of purified DNA from BBa_J04450 (RFP construct) in plasmid backbone pSB1C3.

The first test transformation (1 μl plasmid used, 20 μl plated) showed very poor results:

DNA concentration	0.5pg/μl	5pg/μl	10pg/μl	20pg/μl	50pg/μl
# of colonies	0	0	2	0	13
efficiency (cfu)	0	0	3.8x10⁶	0	3.4x10⁶

Results	efficiency
average	1.4x10⁶
weighted	3.5x10⁶

The second test transformation (3 μl plasmid used, 100 μl plated) showed still poor results but we decided to continue working with our cells:

DNA concentration	0.5pg/μl	5pg/μl	10pg/μl	20pg/μl	50pg/μl
# of colonies	1	13	1	31	28
efficiency (cfu)	2.2x10⁶	2.2x10⁶	1.1x10⁵	1.3x10⁶	4.9x10⁵

Results	efficiency
average	1.3x10⁶
weighted	7.5x10⁵

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