Line 78: | Line 78: | ||
<td><b>Device 1 (J364000)</b></td> | <td><b>Device 1 (J364000)</b></td> | ||
<td bgcolor="#FF0000">J23101(1791)</td> | <td bgcolor="#FF0000">J23101(1791)</td> | ||
− | <td rowspan="3 | + | <td rowspan="3">B0034</td> |
− | <td rowspan=" | + | <td rowspan="7" bgcolor="#40FF00">GFP</td> |
− | <td rowspan=" | + | <td rowspan="7">B0015</td> |
− | <td rowspan=" | + | <td rowspan="8">pSB1C3</td> |
</tr> | </tr> | ||
Line 94: | Line 94: | ||
</tr> | </tr> | ||
+ | <tr> | ||
+ | <td><b>Device 4 (J364003)</b></td> | ||
+ | <td bgcolor="#FF0000">J23101(1791)</td> | ||
+ | <td rowspan="3">BCD2</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td><b>Device 5 (J364004)</b></td> | ||
+ | <td bgcolor="#FA5858">J23106(1185)</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td><b>Device 6 (J364005)</b></td> | ||
+ | <td bgcolor="#FA5858">J23117(162)</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td><b>Positive control (I20270)</b></td> | ||
+ | <td>J23151(N/A)</td> | ||
+ | <td>B0032</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td><b>Negative control (R0040)</b></td> | ||
+ | <td>R0040 (N/A)</td> | ||
+ | <td colspan="3">N/A</td> | ||
+ | </tr> | ||
</table> | </table> | ||
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<h3>What is Bicistronic Device (BCD)?</h3> | <h3>What is Bicistronic Device (BCD)?</h3> | ||
+ | |||
+ | <p><img src="https://static.igem.org/mediawiki/2017/0/08/CuhkBCDinstruction.PNG" style="width:594px;height:216px;"></p> | ||
+ | |||
Bicistronic device (BCD) is a modified ribosome binding site (RBS) with another cistron. The device consists of another cistron (cistron 1) with another RBS (SD2) | Bicistronic device (BCD) is a modified ribosome binding site (RBS) with another cistron. The device consists of another cistron (cistron 1) with another RBS (SD2) | ||
between RBS (SD1) and gene of interest (cistron 2). Also, the stop codon of cistron 1 overlaps the start codon of cistron 2. Ribosome binding efficiency and translation rate will be affected after the secondary structure near the RBS has changed due to the change of gene of interest. This device can maintain the ribosome binding efficiency and translation rate even though the gene of interest has changed. Therefore, it is used to control the amount of fluorescence in this study. BCD is expected to generate a more reliable and precise gene expression. | between RBS (SD1) and gene of interest (cistron 2). Also, the stop codon of cistron 1 overlaps the start codon of cistron 2. Ribosome binding efficiency and translation rate will be affected after the secondary structure near the RBS has changed due to the change of gene of interest. This device can maintain the ribosome binding efficiency and translation rate even though the gene of interest has changed. Therefore, it is used to control the amount of fluorescence in this study. BCD is expected to generate a more reliable and precise gene expression. | ||
+ | |||
+ | |||
+ | <h3>Method</h3> | ||
+ | <p> | ||
+ | We follow exactly the interlab protocol provided by iGEM(link) with the below plate reader setting: | ||
+ | <br> | ||
+ | Microplate used: Corning® clear flat bottom black 96 well plates<br> | ||
+ | Instrument used: BMG LABTECH’s CLARIOstar® | ||
+ | <br> | ||
+ | <b>Instrument Settings for measuring OD600 of LUDOX and Cells:</b> | ||
+ | <br> | ||
+ | <b>- Endpoint settings</b> | ||
</p> | </p> | ||
+ | |||
+ | |||
</div> | </div> | ||
Revision as of 04:45, 8 September 2017