Team:Lethbridge Canada/results

From 2013hs.igem.org

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<p>To detect our preprooxyphysin proteins we did a Silver Stain. The reason for doing a Silver Stain is that it is 10x more sensitive than the standard Coomassie stain. In Lane 2, we did a control of Cbf5-Nop10-Gar1. This is actually three proteins. Because of their re;latively larger size, Cbf5 and Nop10 are found at the top of Lane 2. However, Gar1 is smaller, only 7kDa, and is found to be at the bottom. It is outlined in red. Preprooxyphysin is roughly twice as large as Gar1 at 13kDa and is found to about twice as large as Gar1. It is outlined by orange.</p>
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<p>To detect our prepro-oxyphysin proteins we did a Silver Stain 16.5% Tris-Tricine PAGE. The reason for doing a Silver Stain is that it is 10x more sensitive than the standard Coomassie stain. The tricine in the Tris-TRicine PAGE allows for better and more distinct separation of small proteins in the gel. In Lane 2, we did a control of Cbf5-Nop10-Gar1 proteins. Cbf5 is 43 kDa, Gar1 is 12.3 kDa and Nop10 is only 7.2 kDa. Nop 10 is outlined in red. Prepro-oxyphysin is slightly larger than Nop10 at 10kDa. The bands we suspect are prepro-oxyphysin are outlined by orange.</p>
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<p>We ran a 16.5% tris-tricine PAGE which separates better than than an SDS PAGE. An tris-tricine PAGE contains tricine. Tricine is more effective at separating proteins than the glycine of an SDS PAGE.</p>
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Revision as of 04:58, 21 June 2013

Lane Contents Volume
1 Protein Molecular Weight Marker 20μL
2 Cbf5-Nop10-Gar1 20μL
3 Buffer A wash 30μL
4 Buffer B wash 30μL
5 Buffer E Wash 30μL
6 S-30 s/N 30μL
7 Buffer E – Concentrated Preprooxyphysin 30μL
Gel Gel

To detect our prepro-oxyphysin proteins we did a Silver Stain 16.5% Tris-Tricine PAGE. The reason for doing a Silver Stain is that it is 10x more sensitive than the standard Coomassie stain. The tricine in the Tris-TRicine PAGE allows for better and more distinct separation of small proteins in the gel. In Lane 2, we did a control of Cbf5-Nop10-Gar1 proteins. Cbf5 is 43 kDa, Gar1 is 12.3 kDa and Nop10 is only 7.2 kDa. Nop 10 is outlined in red. Prepro-oxyphysin is slightly larger than Nop10 at 10kDa. The bands we suspect are prepro-oxyphysin are outlined by orange.

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