Team:Bielefeld-Germany/Labjournal/Molecular

From 2013.igem.org

(Difference between revisions)
Line 14: Line 14:
<h1>PCR Protocols and Programs</h1>
<h1>PCR Protocols and Programs</h1>
-
<h3><a name="Phusion">Phusion 25 µl</h3>
+
<h3><a name="cyt_frag1">Phusion 50 µl</h3>
<table border=1 style="float:left">
<table border=1 style="float:left">
Line 20: Line 20:
       <col width="50">
       <col width="50">
       <col width="50">
       <col width="50">
-
<tr> <th>Reagenz</th> <th>1x</th> <th>8x</th> </tr>
+
<tr> <th>Reagent</th>         <th>1x</th> <th>8x</th> </tr>
-
<tr> <td>Buffer 5x</td> <td>5,0</td> <td>44</td> </tr>
+
<tr> <td>Hf-Buffer 5x</td>     <td>10.0</td> <td>88.0</td> </tr>
-
<tr> <td>MgSO4</td> <td>0</td>      <td>0</td> </tr>
+
<tr> <td>dNTPs (10mM)</td>  <td>2.0</td> <td>17.6</td> </tr>
-
<tr> <td>dNTPs</td>  <td>2,5</td> <td>22</td> </tr>
+
<tr> <td>Primer1 (10mM)</td> <td>1.0</td> <td>8,8</td> </tr>
-
<tr> <td>Primer1</td> <td>1,0</td> <td>8,8</td> </tr>
+
<tr> <td>Primer2 (10mM)</td> <td>1.0</td> <td>8,8</td> </tr>
-
<tr> <td>Primer2</td> <td>1,0</td> <td>8,8</td> </tr>
+
<tr> <td>Template (16.5ng)</td> <td>3.0</td> <td>26.4</td> </tr>
-
<tr> <td>Template</td> <td>1,5</td> <td>13,2</td> </tr>
+
<tr> <td>DMSO</td>         <td>4.0</td>    <td>35.2</td> </tr>
-
<tr> <td>DMSO</td> <td>2,0</td>    <td>17,6</td> </tr>
+
<tr> <td>Phusion Polymerase (0.5u)</td>       <td>1.0</td> <td>8.8</td> </tr>
-
<tr> <td>Polymerase</td><td>0,5</td> <td>4,4</td> </tr>
+
<tr> <td>H2O</td>         <td>26.0</td> <td>228.8</td> </tr>
-
<tr> <td>H2O</td>   <td>11,5</td> <td>101,2</td> </tr>
+
</table>
</table>
Line 40: Line 39:
       <col width="50">
       <col width="50">
<tr><th colspan="5">Program Phusion</th></tr>
<tr><th colspan="5">Program Phusion</th></tr>
-
<tr><td>98 °C</td><td>98 °C</td><td>x °C</td><td>72 °C</td><td>72 °C</td></tr>
+
<tr><td>98 °C</td><td>98 °C</td><td>68 °C</td><td>72 °C</td><td>72 °C</td></tr>
-
<tr><td>2 min</td> <td>30 s</td><td>30 s</td> <td>y</td><td>6 min</td></tr>
+
<tr><td>30s</td> <td>10 s</td><td>30 s</td> <td>1:35</td><td>6:00</td></tr>
-
<tr><td colspan="5">x: according to Primer Tm <br> y: 15s/kb
+
<tr><td colspan="5">35 cycles of steps 2 to 4
<br><br>
<br><br>
   <b>Notes</b><br>
   <b>Notes</b><br>
-
   zu beachten ist blablbalbalbalaba <br>
+
   <br>
-
   außerdem ist aufgefallen, dass asdfasdf
+
    
</tr>
</tr>

Revision as of 19:32, 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) 1.0 8.8
H2O 26.0 228.8
Program Phusion
98 °C98 °C68 °C72 °C72 °C
30s 10 s30 s 1:356:00
35 cycles of steps 2 to 4

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
Polymerase0,5 4,4
H2O 11,5 101,2
Program Taq
98 °C98 °Cx °C72 °C72 °C
2 min 30 s30 s y6 min
x: according to Primer Tm
y: 15s/kb

Retrieved from "http://2013.igem.org/Team:Bielefeld-Germany/Labjournal/Molecular"