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Team:Bielefeld-Germany/Labjournal/Molecular
From 2013.igem.org
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| - | <tr> <th>Reagent</th> <th | + | <tr> <th>Reagent</th> <th>1x</th> <th>8x</th> </tr> |
| - | <tr> <td>Hf-Buffer 5x</td> <td>10.0</td> <td>88.0</td> </tr> | + | <tr> <td>Hf-Buffer 5x</td> <td align="right">10.0</td> <td align="right">88.0</td> </tr> |
| - | <tr> <td>dNTPs (10mM)</td> <td>2.0</td> <td>17.6</td> </tr> | + | <tr> <td>dNTPs (10mM)</td> <td align="right">2.0</td> <td align="right">17.6</td> </tr> |
| - | <tr> <td>Primer1 (10mM)</td> <td>1.0</td> <td>8,8</td> </tr> | + | <tr> <td>Primer1 (10mM)</td> <td align="right">1.0</td> <td align="right">8,8</td> </tr> |
| - | <tr> <td>Primer2 (10mM)</td> <td>1.0</td> <td>8,8</td> </tr> | + | <tr> <td>Primer2 (10mM)</td> <td align="right">1.0</td> <td align="right">8,8</td> </tr> |
| - | <tr> <td>Template (16.5ng)</td> <td>3.0</td> <td>26.4</td> </tr> | + | <tr> <td>Template (16.5ng)</td> <td align="right">3.0</td> <td align="right">26.4</td> </tr> |
| - | <tr> <td>DMSO</td> <td>4.0</td> <td>35.2</td> </tr> | + | <tr> <td>DMSO</td> <td align="right">4.0</td> <td align="right">35.2</td> </tr> |
| - | <tr> <td>Phusion Polymerase (0.5u)</td> <td>1.0</td> <td>8.8</td> </tr> | + | <tr> <td>Phusion Polymerase (0.5u)</td> <td> align="right"1.0</td> <td align="right">8.8</td> </tr> |
| - | <tr> <td>H2O</td> <td>26.0</td> <td>228.8</td> </tr> | + | <tr> <td>H2O</td> <td align="right">26.0</td> <td align="right">228.8</td> </tr> |
</table> | </table> | ||
Revision as of 19:41, 30 August 2013
PCR Protocols and Programs
Phusion 50 µl
| Reagent | 1x | 8x |
|---|---|---|
| Hf-Buffer 5x | 10.0 | 88.0 |
| dNTPs (10mM) | 2.0 | 17.6 |
| Primer1 (10mM) | 1.0 | 8,8 |
| Primer2 (10mM) | 1.0 | 8,8 |
| Template (16.5ng) | 3.0 | 26.4 |
| DMSO | 4.0 | 35.2 |
| Phusion Polymerase (0.5u) | align="right"1.0 | 8.8 |
| H2O | 26.0 | 228.8 |
| Program Phusion | ||||
|---|---|---|---|---|
| 98 °C | 98 °C | 68 °C | 72 °C | 72 °C |
| 30s | 10 s | 30 s | 1:35 | 6:00 |
| 35 cycles | ||||
|
Notes:
| ||||
Taq 25 µl
| Reagenz | 1x | 8x |
|---|---|---|
| Buffer 5x | 5,0 | 44 |
| MgSO4 | 0 | 0 |
| dNTPs | 2,5 | 22 |
| Primer1 | 1,0 | 8,8 |
| Primer2 | 1,0 | 8,8 |
| Template | 1,5 | 13,2 |
| DMSO | 2,0 | 17,6 |
| Polymerase | 0,5 | 4,4 |
| H2O | 11,5 | 101,2 |
| Program Taq | ||||
|---|---|---|---|---|
| 98 °C | 98 °C | x °C | 72 °C | 72 °C |
| 2 min | 30 s | 30 s | y | 6 min |
| x: according to Primer Tm y: 15s/kb | ||||
