Team:University College London/LabBook/Week14

From 2012.igem.org

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'''Aim:''' To characterise nuclease activity  
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'''Aim:''' To characterise nuclease activity BL21 cells
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'''Conclusion:''' works bust in comparision to one found naturally in wnu it produces less nucles, as expected.
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'''Conclusion:'''  
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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.

Revision as of 00:31, 27 September 2012

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: http://2012.igem.org/Team:University_College_London/Protocols/Competency2

Tuesday 11.09.12

Aim: Day 3 of Generating competent cells


Method: http://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.