Team:Freiburg/Results

From 2011.igem.org

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(Plastic binding domain)
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In case of the pbd-bound GFP it is striking that after the solution has already been diluted by one washing step, the percentage of protein that can be washed away diminishes. It comes to mind that the massive lost of pbd-tagged GFP after the first washing step might be due to an oversaturation of pbd-GFP in the solution.  In this case there would be much more pbd-GFP than place on the plastic surface so that most of the protein could be eluated.
In case of the pbd-bound GFP it is striking that after the solution has already been diluted by one washing step, the percentage of protein that can be washed away diminishes. It comes to mind that the massive lost of pbd-tagged GFP after the first washing step might be due to an oversaturation of pbd-GFP in the solution.  In this case there would be much more pbd-GFP than place on the plastic surface so that most of the protein could be eluated.
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As shown in picture 2 the percentage of eluted GFP diminishes when the used start concentration is lower. As the used [[Image: pbd_better_than_GFP.jpg|250px|right| thumb | Picture 3: if the start concentration is not oversaturated pbd-coupled is much more resistant to washing steps than GFP alone.]]  
As shown in picture 2 the percentage of eluted GFP diminishes when the used start concentration is lower. As the used [[Image: pbd_better_than_GFP.jpg|250px|right| thumb | Picture 3: if the start concentration is not oversaturated pbd-coupled is much more resistant to washing steps than GFP alone.]]  
To investigate this phenomenon  we compared different start concentrations of pbd-GFP concerning the amount of pbd-GFP that could be washed away.
To investigate this phenomenon  we compared different start concentrations of pbd-GFP concerning the amount of pbd-GFP that could be washed away.
polystyrene micro titer plates provide only a limited surface for the pbd to bind, the solution shouldn’t be oversaturated with plastic binding protein.
polystyrene micro titer plates provide only a limited surface for the pbd to bind, the solution shouldn’t be oversaturated with plastic binding protein.
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In a range of 1-30ng/µL start concentration there remains about seven times more pbd-GFP after washing than "normal" GFP (see picture 3).  
In a range of 1-30ng/µL start concentration there remains about seven times more pbd-GFP after washing than "normal" GFP (see picture 3).  

Revision as of 01:50, 22 September 2011


This is the wiki page
of the Freiburger student
team competing for iGEM 2011.
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