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| | <div class="story"> | | <div class="story"> |
| | <div class="paragraphs"> | | <div class="paragraphs"> |
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| | + | <h2>Our Favorite Composite Part: <a href="http://parts.igem.org/Part:BBa_K2751013">BBa_K2751013</a></h2> |
| | + | <h2>pCMV-ALB-mEGFP internal control</h2> |
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| | + | <h3>Usage and Biology</h3> |
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| | + | <p>This part contains the pCMV-ALB-mEGFP sequence, and is used as our internal control plasmid. The CMV promoter allows mEGFP to be expressed constantly when transfected into cells. Moreover, the signal peptide (ALB) allows mEGFP to be secreted extracellularly, so that signal detection can become much easier. </p> |
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| | + | <img src="https://static.igem.org/mediawiki/parts/b/b8/T--NYMU-Taipei--IC1.png" style="width:900px;"> |
| | + | <img src="https://static.igem.org/mediawiki/parts/0/09/T--NYMU-Taipei--IC2.png" style="width:500px;"> |
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| | + | |
| | + | <h3>Characterization</h3> |
| | + | <p>We ran electrophoresis to check if the length of the part is correct. The length of CMV promoter, ALB sequence, and mEGFP sequence are 290bp, 56bp, and 720bp respectively. Therefore, we expect a product of 1066bp (1099bp along with a biobrick prefix). Gel photo is shown in the figure below.</p> |
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| | + | <img src="https://static.igem.org/mediawiki/parts/4/46/T--NYMU-Taipei--parts-IC3.png" style="width:250px;"> |
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| | + | <h3>Functionality Test</h3> |
| | + | <p>We transfected HEK293T cells via the GenJet Plus DNA transfection reagent and then collected cell supernatant for fluorescence measurement after 24 and 48 hours. Transparent alpha-MEM medium was used to minimize background noise. We used the Varioskan LUX spectral scanning multimode reader to measure fluorescence levels. Excitation wavelength was 488nm, emission wavelength was 509nm, and bandwidth was 5nm. Result is shown in the figure below. Blank medium control is taken from fresh medium, and cultured cell control is taken from the supernatant of cells that have not been transfected with the internal control plasmid. </p> |
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| | + | <img src="https://static.igem.org/mediawiki/parts/8/8d/T--NYMU-Taipei--parts-IC4.png" style="width:500px;"> |
| | + | |
| | + | </div> |
| | + | </div> |
| | + | |
| | + | <div class="story"> |
| | + | <div class="paragraphs"> |
| | + | |
| | + | <h2>List of Composite Parts</h2> |
| | | | |
| | <table style="width:100%"> | | <table style="width:100%"> |
| − | <tr>
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| − | <td><a href="http://parts.igem.org/Part:BBa_K2751013">BBa_K2751013</a></td>
| + | |
| − | <td>pCMV-ALB-mEGFP internal control</td>
| + | |
| − | <td>Composite</td>
| + | |
| − | <td>1066</td>
| + | |
| − | </tr>
| + | |
| | <tr> | | <tr> |
| | <th>Part Number</th> | | <th>Part Number</th> |
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| | <th>Type</th> | | <th>Type</th> |
| | <th>Length(bp)</th> | | <th>Length(bp)</th> |
| | + | </tr> |
| | + | <tr> |
| | + | <td><a href="http://parts.igem.org/Part:BBa_K2751013">BBa_K2751013</a></td> |
| | + | <td>pCMV-ALB-mEGFP internal control</td> |
| | + | <td>Composite</td> |
| | + | <td>1066</td> |
| | </tr> | | </tr> |
| | <tr> | | <tr> |
