Team:Bielefeld-Germany/Labjournal/Molecular

(Difference between revisions)

Revision as of 11:30, 6 September 2013

*a2 **b2 ***c2

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:

@@ Line 59: / Line 59: @@
 </div>
 <div class="standard_protocol">
-<h2>Phusion PCR</h2>
 <table border=1 style="float:left; padding-left:10px">
        <col width="150">
@@ Line 75: / Line 74: @@
 </table>
-<br><br><br><br><br><br><br><br><br><br>
-<h2>Colony PCR</h2>
-<p> - Pick one colony with a sterile tip and elute it in 100 µL ddH20 or medium </p>
-<p> - Store the colony in 4 °C while colony PCR is running <p>
-<p> - One reaction mix contains: </p>
-<p> - 10 µL 5x buffer </p>
-<p> - 2 µL MgCl2 (25 mM stock) </p>
-<p> - 1 µL dNTPs </p>
-<p> - 0.5 µL primer mix (prefix/suffix primers or sequencing primers) </p>
-<p> - 35.25 µL ddH2O </p>
-<p> - 0.25 µL  BioTaq polymerase (BIOLINE) </p>
-<p> - 1 µL template </p>
-<p> - PCR program: </p>
-<p>     - Start: 3 min, 98 °C </p>
-<p>     - 30 cycles of: </p>
-<p>        - 30 s, 98 °C </p>
-<p>        - 30 s, 55 °C </p>
-<p>        - 30 s / 1 kb template, 72 °C </p>
-<p>     - Finish: 5 min, 72 °C </p>
-<p> Gel electrophoresis: check the fragment size </p>
-<p> Plate the correct colony </p>
-<br><br><br><br><br><br><br><br>
+<br><br><br><br><br><br><br>
 </div>
+*a2
+**b2
+***c2
+<br><br>
 <div class="cyt_frag1_headline">
 <a name="cyt_frag1"><span style="color:#ff6600">[Cytochromes]</span> Phusion 50ul for mtrCAB Fragment1</span></a>