Difference between revisions of "Team:Warwick/Project"
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| − | <p> | + | <p><p style="float: right;"><img src="https://static.igem.org/mediawiki/2015/3/31/Warwickoverview2.png" height="200px" width="400px" border="50px"></p> |
| − | As a starting point we are conducting tests for the expression of the fusion protein, and working on a method of placing DNA on glass slides. Then we can test the binding of DNA to the slides in simple patterns to view the system. Currently we have demonstrated we can adhere oligonucleotides to the glass slide as shown in Fig.2. Fluorescent oligonucleotides (with amine groups) were introduced to 2 glass slides prepared; one treated with GOPTS, the other only cleaned. Both slides were washed and inspected with a FITC green filter. In theory only the treated slide should show fluorescence, and the results were as expected. | + | As a starting point we are conducting tests for the expression of the fusion protein, and working on a method of placing DNA on glass slides. Then we can test the binding of DNA to the slides in simple patterns to view the system. Currently we have demonstrated we can adhere oligonucleotides to the glass slide as shown in Fig.2. Fluorescent oligonucleotides (with amine groups) were introduced to 2 glass slides prepared; one treated with GOPTS, the other only cleaned. Both slides were washed and inspected with a FITC green filter. In theory only the treated slide should show fluorescence, and the results were as expected. |
| + | <p style="float: right;"><img src="https://static.igem.org/mediawiki/2015/3/3a/Warwickoverview3.png" height="200px" width="400px" border="50px"></p> | ||
<br>To demonstrate that the E.coli receiving our part construct are being expressed we added a flag tag to the part inserted, allowing us to treat cells with an anti-FLAG antibody and visualise using fluorescent microscopy. Wild-type, transformed and uninduced, and transformed and induced DH5α Z1 cells were dried onto cover slips and fixed. The cells were then treated with an anti-FLAG antibody, followed by an anti-mouse fluorescent antibody. The slides were then examined with a FITC green filter shown in Fig.3. Although random flourescence is seen in A this is likely due to poor antibody removal based on the areas of flourescence. The brightest floursecnce and clear cellular outlines are seen in C in the induced DH5α Z1 cells. | <br>To demonstrate that the E.coli receiving our part construct are being expressed we added a flag tag to the part inserted, allowing us to treat cells with an anti-FLAG antibody and visualise using fluorescent microscopy. Wild-type, transformed and uninduced, and transformed and induced DH5α Z1 cells were dried onto cover slips and fixed. The cells were then treated with an anti-FLAG antibody, followed by an anti-mouse fluorescent antibody. The slides were then examined with a FITC green filter shown in Fig.3. Although random flourescence is seen in A this is likely due to poor antibody removal based on the areas of flourescence. The brightest floursecnce and clear cellular outlines are seen in C in the induced DH5α Z1 cells. | ||
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Revision as of 16:11, 17 September 2015
