Team:Macquarie Australia/Protocols/PCR

From 2013.igem.org

(Difference between revisions)
Line 24: Line 24:
     Initial denaturation 98°C for 30 secs<br>
     Initial denaturation 98°C for 30 secs<br>
     <b>Followed by 30 repeats of:</b><br>
     <b>Followed by 30 repeats of:</b><br>
-
         Denaturation - 98 °C @ 10 secs<br>
+
         <b>Denaturation -</b> 98 °C for 10 secs<br>
-
         Annealing - 60°C      @ 10 secs<br>
+
         <b>Annealing -</b> 60°C      for 10 secs<br>
-
         Extension - 72°C        @ 2 mins<br>
+
         <b>Extension -</b> 72°C        for 2 mins<br>
-
         Final extension - 72°C  @ 10 mins<br>
+
         <b>Final extension -</b> 72°C  for 10 mins<br>

Revision as of 06:51, 13 September 2013


Polymerase Chain Reaction


PCR mixture

4.0ul 5x phusion buffer
0.4ul dNTPs
0.6ul DMSO
0.2ul Polymerase
11.8ul water
Total 17ul
1ul forward primer
1ul reverse primer
1ul template


PCR settings

Initial denaturation 98°C for 30 secs
Followed by 30 repeats of:
Denaturation - 98 °C for 10 secs
Annealing - 60°C for 10 secs
Extension - 72°C for 2 mins
Final extension - 72°C for 10 mins

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