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Team:Bielefeld-Germany/Labjournal/Molecular
From 2013.igem.org
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<h1>PCR Protocols and Programs</h1> | <h1>PCR Protocols and Programs</h1> | ||
| + | |||
| + | |||
| + | <div class="standard_headline"> | ||
| + | <a name="standard_protocol"><span style="color:#ff6600">[Standard Protocols]</span> Standard protocols</span></a> | ||
| + | </div> | ||
| + | <div class="standard_protocol"> | ||
| + | <table border=1 style="float:left; padding-left:10px"> | ||
| + | <col width="150"> | ||
| + | <col width="50"> | ||
| + | <col width="50"> | ||
| + | <tr> <th>Reagent</th> <th>1x</th> <th>8x</th> </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 align="right">2.0</td> <td align="right">17.6</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 align="right">1.0</td> <td align="right">8.8</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 align="right">4.0</td> <td align="right">35.2</td> </tr> | ||
| + | <tr> <td>Phusion (0.5u)</td> <td align="right">1.0</td> <td align="right">8.8</td> </tr> | ||
| + | <tr> <td>H2O</td> <td align="right">26.0</td> <td align="right">228.8</td> </tr> | ||
| + | |||
| + | </table> | ||
| + | |||
| + | <br><br><br><br><br><br><br> | ||
| + | </div> | ||
| + | |||
| + | |||
<div class="cyt_frag1_headline"> | <div class="cyt_frag1_headline"> | ||
<a name="cyt_frag1"><span style="color:#ff6600">[Cytochromes]</span> Phusion 50ul for mtrCAB Fragment1</span></a> | <a name="cyt_frag1"><span style="color:#ff6600">[Cytochromes]</span> Phusion 50ul for mtrCAB Fragment1</span></a> | ||
Revision as of 10:11, 6 September 2013
PCR Protocols and Programs
| 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 (0.5u) | 1.0 | 8.8 |
| H2O | 26.0 | 228.8 |
| 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 (0.5u) | 1.0 | 8.8 |
| H2O | 26.0 | 228.8 |
| Program Phusion | ||||
|---|---|---|---|---|
| 98 °C | 98 °C | 68 °C | 72 °C | 72 °C |
| 30s | 10s | 30s | 1:35 | 6:00 |
| 35 cycles | ||||
|
Notes:
Size: 1800bp Primer1: Primer2: | ||||
| Reagent | 1x | 8x |
|---|---|---|
| Hf-Buffer 5x | 10.0 | 88.0 |
| dNTPs (10mM) | 2.5 | 22.0 |
| Primer1 (10mM) | 1.0 | 8.8 |
| Primer2 (10mM) | 1.0 | 8.8 |
| Template (5.5ng/ul) | 2.0 | 17.6 |
| DMSO | 3.0 | 26.4 |
| Phusion (0.5u) | 1.0 | 8.8 |
| H2O | 29.5 | 259.6 |
| Program Phusion | ||||
|---|---|---|---|---|
| 98 °C | 98 °C | 60 °C | 72 °C | 72 °C |
| 1:00 | 10s | 30s | 30s | 6:00 |
| 35 cycles | ||||
|
Notes:
Size: 330bp Primer1: Primer2: | ||||
| Reagent | 1x | 8x |
|---|---|---|
| Hf-Buffer 5x | 10.0 | 88.0 |
| dNTPs (10mM) | 2.5 | 22.0 |
| Primer1 (10mM) | 1.0 | 8.8 |
| Primer2 (10mM) | 1.0 | 8.8 |
| Template (5.5ng/ul) | 2.0 | 17.6 |
| DMSO | 3.0 | 26.4 |
| Phusion (0.5u) | 1.0 | 8.8 |
| H2O | 29.5 | 259.6 |
| Program Phusion | ||||
|---|---|---|---|---|
| 98 °C | 98 °C | 70 °C | 72 °C | 72 °C |
| 30s | 10s | 30s | 50s | 6:00 |
| 35 cycles | ||||
|
Notes:
Size: 3000bp Primer1: Primer2: | ||||
| Reagent | 1x | 8x |
|---|---|---|
| GC-Buffer 5x | 10.0 | 88.0 |
| dNTPs (10mM) | 2.0 | 17.6 |
| Primer1 (10mM) | 1.25 | 11.0 |
| Primer2 (10mM) | 1.25 | 11.0 |
| Template (5.5ng/ul) | 1.0 | 8.8 |
| DMSO | 1.5 | 13.2 |
| Phusion (0.5u) | 0.5 | 4.4 |
| H2O | 32.5 | 286.0 |
| Program Phusion | ||||
|---|---|---|---|---|
| 98 °C | 98 °C | 70 °C | 72 °C | 72 °C |
| 45s | 30s | 1:00 | 45s | 5:00 |
| 35 cycles | ||||
|
Notes:
Size: 2070bp Primer1_Gibson: Primer2_Gibson: Primer1: Primer2: | ||||
| Reagent | 1x | 8x |
|---|---|---|
| Hf-Buffer 5x | 10.0 | 88.0 |
| dNTPs (10mM) | 2.5 | 22.0 |
| Primer1 (10mM) | 1.0 | 8.8 |
| Primer2 (10mM) | 1.0 | 8.8 |
| Template (5.5ng/ul) | 2.0 | 17.6 |
| DMSO | 3.0 | 26.4 |
| Phusion (0.5u) | 1.0 | 8.8 |
| H2O | 29.5 | 259.6 |
| Program Phusion | ||||
|---|---|---|---|---|
| 98 °C | 98 °C | 60 °C | 72 °C | 72 °C |
| 30s | 10s | 30s | 1:15 | 6:15 |
| 35 cycles | ||||
|
Notes:
Size: 5150bp Primer1: Primer2: | ||||
| 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 (5.5ng/ul) | 3.0 | 26.4 |
| DMSO | 3.0 | 26.4 |
| Phusion (0.5u) | 1.0 | 8.8 |
| H2O | 29.5 | 255.2 |
| Program Phusion | ||||
|---|---|---|---|---|
| 98 °C | 98 °C | 68 °C | 72 °C | 72 °C |
| 30s | 10s | 30s | 1:35 | 6:00 |
| 35 cycles | ||||
|
Notes:
Size: 6311bp Primer1: Primer2: | ||||
