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User:University of Washington/25 July 2008
From 2008.igem.org
(Difference between revisions)
(→LuxR from AraC and TetR) |
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</table> | </table> | ||
*Thermocycling | *Thermocycling | ||
| - | <table | + | <table> |
<tr> | <tr> | ||
<th>Segment</th><th>Cycles</th><th>Temperature</th><th>Time</th> | <th>Segment</th><th>Cycles</th><th>Temperature</th><th>Time</th> | ||
</tr> | </tr> | ||
| - | <tr> | + | <tr align=center> |
<td>1</td><td>1</td><td>95°C</td><td>1 minute</td> | <td>1</td><td>1</td><td>95°C</td><td>1 minute</td> | ||
</tr> | </tr> | ||
| - | <tr> | + | <tr align=center> |
<td rowspan=3>2</td><td rowspan=3>18</td><td>95°C</td><td>50 seconds</td> | <td rowspan=3>2</td><td rowspan=3>18</td><td>95°C</td><td>50 seconds</td> | ||
</tr> | </tr> | ||
| - | <tr> | + | <tr align=center> |
<td>60°C</td><td>50 seconds</td> | <td>60°C</td><td>50 seconds</td> | ||
</tr> | </tr> | ||
| - | <tr> | + | <tr align=center> |
<td>68°C</td><td>1 minute/kb of plasmid length</td> | <td>68°C</td><td>1 minute/kb of plasmid length</td> | ||
</tr> | </tr> | ||
| - | <tr> | + | <tr align=center> |
<td>3</td><td>1</td><td>68°C</td><td>7 minutes</td> | <td>3</td><td>1</td><td>68°C</td><td>7 minutes</td> | ||
</tr> | </tr> | ||
Revision as of 21:26, 25 July 2008
LuxR from AraC and TetR
-Received primers to biobrick Elowitz's plasmid.
-Did PCR amplification on Elowitz's plasmid.
- Reaction Set-Up
| reagents | amount per reaction(ul) |
|---|---|
| sterile dH2O | 34.5 |
| buffer for Taq (-MgCl2) | 5 |
| 10 mM dNTPs | 1 |
| MgCl2 | 2 |
| F primer(#436) | 2.5 |
| R primer(#436) | 2.5 |
| Taq | 0.5 |
| DNA template | 2 |
- Thermocycling
| Segment | Cycles | Temperature | Time |
|---|---|---|---|
| 1 | 1 | 95°C | 1 minute |
| 2 | 18 | 95°C | 50 seconds |
| 60°C | 50 seconds | ||
| 68°C | 1 minute/kb of plasmid length | ||
| 3 | 1 | 68°C | 7 minutes |
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