Team:SDU-Denmark/protocols

From 2010.igem.org

(Difference between revisions)
(CP1.2)
(CP1.1)
Line 29: Line 29:
Premix TAQ-PCR (no proofreading): <br>
Premix TAQ-PCR (no proofreading): <br>
For 1 PCR reaction <br><br>
For 1 PCR reaction <br><br>
-
2.5 µL 10x TAQ-buffer + MgCl2 <br><br>
+
• 5 µL 10x TAQ-buffer<br><br>
-
0.5 µL 10mM dNTP mix <br><br>
+
• 2 µL MgCl2 <br><br>
-
1.25 µL 10pmol/µL forward primer <br><br>
+
1 µL 10mM dNTP mix <br><br>
-
1.25 µL 10pmol/µL reverse primer <br><br>
+
2 µL 10pmol/µL forward primer <br><br>
-
0.25 µL TAQ polymerase enzyme (add just before PCR run)<br><br>
+
2 µL 10pmol/µL reverse primer <br><br>
-
• 19.25 µL H2O <br><br>
+
35 µL Water<br><br><br>
-
Total vol.: 25 µL <br><br>
+
• 1 µL DNA <br><br>
 +
• 1 µL TAQ polymerase enzyme '''(add just before PCR run)'''<br><br>
 +
Total vol.: 49 µL <br><br>
The TAQ polymerase has no proofreading, and should therefore only be used for size determination of DNA fragments. When PCR-product is to be purified and used for further experiments always use Phu polymerase!!!
The TAQ polymerase has no proofreading, and should therefore only be used for size determination of DNA fragments. When PCR-product is to be purified and used for further experiments always use Phu polymerase!!!
Make enough premix for your number of colonies +3. <br><br>
Make enough premix for your number of colonies +3. <br><br>

Revision as of 14:41, 15 July 2010