Team:Goettingen/Results
Project Results
Transformation Efficiency Kit, RFP construct (iGEM)
Before using our competent E. coli TOP10 cells in the important experiments, we used the Transformation Efficiency 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 (50 μl of competent cells, 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 (200 μl of competent cells, 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 |
35 |
28 |
|
efficiency (cfu) |
8.0x106 |
1.0x107 |
4.0x105 |
7.0x106 |
2.3x106 |
|
Results |
efficiency |
|
average |
5.6x106 |
|
weighted |
3.7x106 |
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