Difference between revisions of "Team:Edinburgh/Characterisation Part"
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We improved and expanded characterisation on the part BBa_K1321357, a carbohydrate binding domain (CBD) submitted by the Imperial College 2014 team. After confirming expression through SDS PAGE we incubated iso-standard paper chads from Whatman 54 filter paper in a crude cell lysate containing the CBD bound to super folded GFP for ten minutes. We then let the protein saturated chads sit in a large volume of 1 x PBS buffer at pH 7.4 (to remain consistent with imperial’s initial characterisation.) At different time points chads were taken from the buffer and placed into a 96 well plate, over 30 minutes. The decrease in fluorescence over time was indicative of relative binding affinity, to Whatman 54 paper. The graph below shows our results. | We improved and expanded characterisation on the part BBa_K1321357, a carbohydrate binding domain (CBD) submitted by the Imperial College 2014 team. After confirming expression through SDS PAGE we incubated iso-standard paper chads from Whatman 54 filter paper in a crude cell lysate containing the CBD bound to super folded GFP for ten minutes. We then let the protein saturated chads sit in a large volume of 1 x PBS buffer at pH 7.4 (to remain consistent with imperial’s initial characterisation.) At different time points chads were taken from the buffer and placed into a 96 well plate, over 30 minutes. The decrease in fluorescence over time was indicative of relative binding affinity, to Whatman 54 paper. The graph below shows our results. | ||
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Revision as of 03:30, 19 September 2015
We improved and expanded characterisation on the part BBa_K1321357, a carbohydrate binding domain (CBD) submitted by the Imperial College 2014 team. After confirming expression through SDS PAGE we incubated iso-standard paper chads from Whatman 54 filter paper in a crude cell lysate containing the CBD bound to super folded GFP for ten minutes. We then let the protein saturated chads sit in a large volume of 1 x PBS buffer at pH 7.4 (to remain consistent with imperial’s initial characterisation.) At different time points chads were taken from the buffer and placed into a 96 well plate, over 30 minutes. The decrease in fluorescence over time was indicative of relative binding affinity, to Whatman 54 paper. The graph below shows our results.
![](https://static.igem.org/mediawiki/2015/0/07/5.20M.jpeg)
We improved and expanded characterisation on the part BBa_K1321356, a carbohydrate binding domain (CBD) submitted by the Imperial College 2014 team. After confirming expression through SDS PAGE we incubated iso-standard paper chads from Whatman 54 filter paper in a crude cell lysate containing the CBD bound to super folded GFP for ten minutes. We then let the protein saturated chads sit in a large volume of 1 x PBS buffer at pH 7.4 (to remain consistent with imperial’s initial characterisation.) At different time points 5 chads were taken from the buffer and placed into a 96 well plate, over 30 minutes. The decrease in fluorescence over time was indicative of relative binding affinity, to Whatman 54 paper. The graph below shows our results.