Team:EPF-Lausanne/Protocols/Klenow

From 2009.igem.org

(Difference between revisions)
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{{EPF-Lausanne09}}
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<div CLASS="epfltrick">__TOC__
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</div><div CLASS="epfl09">
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<html>
<html>
-
<head>
+
<body>
 +
<form action="input_button.htm">
 +
<p align="right">
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<input type="button" name="lien" value="Back to Protocols"
 +
onClick="self.location.href='https://2009.igem.org/Team:EPF-Lausanne/Protocols'">
 +
</p>
 +
</form>
 +
</body>
 +
</html>
 +
<html><center>
 +
<font size="6" color="#007CBC">1.5 Steps PCR Protocol</font>
 +
</center></html>
 +
<br>
 +
----
 +
<br>
 +
<br>
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<style>
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'''1.''' Do the following mixing:
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/*prop. de la premiere ligne*/
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-
.firstHeading {
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-
  display:none;
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-
}
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.top-section {
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-
  display:none;
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-
}
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/*box igem en bas*/
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<center>
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#footer-box {
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{| class="wikitable" style="text-align:left; width:30%;"
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  float: bottom;
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|-
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}
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! scope=row | dNTP
 +
|1 μl
 +
|-
 +
! scope=row | 5x Buffer + MgCl2
 +
|10 μl
 +
|-
 +
! scope=row | Primer Mix
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|1 μl
 +
|-
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! scope=row | template
 +
|0.5 μl
 +
|-
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! scope=row | HiFi Plus DNAp
 +
|0.5 μl
 +
|-
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! scope=row | dH2O
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|36.5 μl
 +
|-
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! scope=row | '''Final Volume'''
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|'''50 μl'''
 +
|}
 +
</center>
-
/*couleur du box ou on peut printer*/
 
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#content {
 
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          /*background:white url("https://static.igem.org/mediawiki/2009/d/d6/Fond_igem.jpg") fixed;*/
 
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          border-width: 0px;
 
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}
 
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/*couleur du sous-box a l'intérieur de content, garder la couleur a neutre*/
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'''2.''' Make it run '''10''' cycles:
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<center>
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          padding: 0px;
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{| class="wikitable" style="text-align:left; width:20%;"
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          margin: 0px;
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|-
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! scope=row | 1&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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|94°C
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|4:00
 +
|-
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! scope=row | 2&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
|94°C
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|0:30
 +
|-
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! scope=row | 3&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
|55°C
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|1:00
 +
|-
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! scope=row | 4&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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|72°C
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|2:00
 +
|-
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! scope=row | 5&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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|Cycle 2-4 10 times
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|
 +
|-
 +
! scope=row | 6&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
|72°C
 +
|7:00
 +
|-
 +
! scope=row | 7&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
|4°C
 +
|&infin;
 +
|}
 +
</center>
-
}
 
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/*------------------------calendar-----------------------*/
 
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.heading {
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'''3.''' We now add the standard iGEM primers which finish the extension, and do the following cycles:
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background-color: #007CBC;
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<center>
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font-size: 20px;
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|-
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! scope=row | 1&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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|94°C
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background-color: #E5E9FF;
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|4:00
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font-size: 16px;
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|-
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color: #007CBC;
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! scope=row | 2&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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|94°C
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|0:30
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! scope=row | 3&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
|55°C
 +
|1:00
 +
|-
 +
! scope=row | 4&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
|72°C
 +
|2:00
 +
|-
 +
! scope=row | 5&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
|Cycle 2-4 25-30 times
 +
|
 +
|-
 +
! scope=row | 6&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
|72°C
 +
|7:00
 +
|-
 +
! scope=row | 7&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
|4°C
 +
|&infin;
 +
|}
 +
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Revision as of 19:39, 20 October 2009



1.5 Steps PCR Protocol



1. Do the following mixing:

dNTP 1 μl
5x Buffer + MgCl2 10 μl
Primer Mix 1 μl
template 0.5 μl
HiFi Plus DNAp 0.5 μl
dH2O 36.5 μl
Final Volume 50 μl


2. Make it run 10 cycles:

1             94°C 4:00
2             94°C 0:30
3             55°C 1:00
4             72°C 2:00
5             Cycle 2-4 10 times
6             72°C 7:00
7             4°C ∞


3. We now add the standard iGEM primers which finish the extension, and do the following cycles:

1             94°C 4:00
2             94°C 0:30
3             55°C 1:00
4             72°C 2:00
5             Cycle 2-4 25-30 times
6             72°C 7:00
7             4°C ∞


Retrieved from "http://2009.igem.org/Team:EPF-Lausanne/Protocols/Klenow"