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Team:University College London/LabBook/Week14
From 2012.igem.org
Contents |
Monday 10.09.12
Aim: Day 2 of Generating competent cells. The aim is to incubate cells in the presence of CaCl2 to prepare the cell wall, such that it becomes permeable to DNA
Method: https://2012.igem.org/Team:University_College_London/Protocols/Competency2
Tuesday 11.09.12
Aim: Day 3 of Generating competent cells
Method: https://2012.igem.org/Team:University_College_London/Protocols/Competency3
14-2
14-3
14-4
Friday 14.09.13
Aim: To characterise nuclease activity BL21 cells
Method:
To add protocol
Results:
The following table shows readings of the diameters and OD at different time points:
| Date | Time | Colony diameter/mm | Halo diameter/mm | Absorbance at 600 OD |
|---|---|---|---|---|
| Friday | 12.30 | 0 | 0 | 0 |
| Friday | 16.30 | 0 | 0 | 0 |
| Friday | 19.30 | 0 | 0 | 0 |
| Friday | 22.30 | 0 | 0 | 0 |
| Saturday | 01.30 | 0.5 | 1.5 | 0.068 |
| Saturday | 04.30 | 1 | 2 | 0.098 |
| Saturday | 07.30 | 1.5 | 3 | 0.159 |
| Saturday | 10.30 | 1.5 | 3.5 | 0.192 |
| Saturday | 12.30pm | 2 | 4 | 0.203 |
| Saturday | 14.30 | 2 | 4 | 0.209 |
| Saturday | 16.30 | 2.5 | 5 | 0.215 |
The average depth of the colonies was noted to be 0.5 - 1mm
Conclusion:
It can be seen that nuclease works as expected in BL21 cells. However, nuclease activity is less efficient when compared to nuclease activity in WnU cells. This is in line with expectations, since nuclease is found naturally in WnU cells but not in BL21 cells.
