Difference between revisions of "Team:Tufts/Results"
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The genes were then ligated into vectors and transformed into JM-109 E. Coli bacteria cells in order to replicate the vectors. The bacteria were left to grow overnight, and then the plasmid was isolated from them. | The genes were then ligated into vectors and transformed into JM-109 E. Coli bacteria cells in order to replicate the vectors. The bacteria were left to grow overnight, and then the plasmid was isolated from them. | ||
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− | |||
− | |||
− | |||
− | |||
pHsp70-Cas9 | pHsp70-Cas9 | ||
− | Sample | + | <table> |
− | Concentration (ng/μL) | + | <tr> |
− | A | + | <td>Sample</td> |
− | 45.7 | + | <td>Concentration (ng/μL)</td> |
− | B | + | </tr> |
− | 46.3 | + | <tr> |
− | C | + | <td>A</td> |
− | 40.4 | + | <td>45.7</td> |
+ | </tr> | ||
+ | <tr> | ||
+ | <td>B</td> | ||
+ | <td>46.3</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>C</td> | ||
+ | <td>40.4</td> | ||
+ | </tr> | ||
+ | </table> | ||
Finally, the Cas9 gene was digested, purified, and ligated upstream of the aTcdB gene in the pHis1522 plasmid. The results were confirmed on a protein gel. | Finally, the Cas9 gene was digested, purified, and ligated upstream of the aTcdB gene in the pHis1522 plasmid. The results were confirmed on a protein gel. | ||
NanoDrop of pHis1522-aTcdB-Cas9 Results | NanoDrop of pHis1522-aTcdB-Cas9 Results | ||
− | Sample | + | |
− | Concentration (ng/μL) | + | <table> |
− | 1 | + | <tr> |
− | 49.9 | + | <td>Sample</td> |
− | 2 | + | <td>Concentration (ng/μL)</td> |
− | 72.1 | + | </tr> |
− | 3 | + | <tr> |
− | 1702.6 | + | <td>1</td> |
− | 4 | + | <td>49.9</td> |
− | 426.6 | + | </tr> |
− | 5 | + | <tr> |
− | 42.7 | + | <td>2</td> |
+ | <td>72.1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>3</td> | ||
+ | <td>1702.6</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>4</td> | ||
+ | <td>426.6</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>5</td> | ||
+ | <td>42.7</td> | ||
+ | </tr> | ||
+ | </table> | ||
The gel of samples 1-5.A faint band (not clearly visible in image) at ~4.2 kb in 3, indicates Cas9 presence/success. | The gel of samples 1-5.A faint band (not clearly visible in image) at ~4.2 kb in 3, indicates Cas9 presence/success. |
Revision as of 23:33, 18 September 2015
The first few steps of the project involved purifying and checking both of the genes for Cas9 and aTcdB. This involved purifying the products of PCR and running them on electrophoresis gels, as well as using a nanodrop machine to identify the concentration and purity of DNA recovered.
1. pHis1522-aTcdBSample | Concentration (ng/μL) |
A | 135.5 |
B | 170.2 |
Sample | Concentration (ng/μL) |
A | 32.9 |
B | 33.3 |
C | 118.3 |
![](https://static.igem.org/mediawiki/2015/3/3e/Tuftsresultsgel1.png)
Sample | Concentration (ng/μL) |
A | 45.7 |
B | 46.3 |
C | 40.4 |
Sample | Concentration (ng/μL) |
1 | 49.9 |
2 | 72.1 |
3 | 1702.6 |
4 | 426.6 |
5 | 42.7 |
![](https://static.igem.org/mediawiki/2015/8/86/Tuftsresultsgel2.png)