PCR

From 2011.igem.org

(Difference between revisions)
Line 37: Line 37:
72°C - 3.00 min
72°C - 3.00 min
-
repeat 3-5 for 40 cycles
+
repeat 3-5 for 35 cycles
72°C - 10.00 min
72°C - 10.00 min

Revision as of 02:45, 29 September 2011

Materials:

DNA template

Invitrogen platinium blue supermix

Forward primer

Reverse Primer

Molecular grade H2O

Procedure:

1. prepare a stock solution per PCR reaction to be done of

-12.5ul PCR supermix

-2.5ul Forward primer

-2.5ul Reverse primer

-5.5ul molecular grade H2O

2. aliquot into 0.6ml PCR tubes for each PCR reaction to be done

3. add 2ul of DNA template

4. run PCR program

1. 94°C - 5.00 min

2. 94°C - 30 sec

51°C - 1.00 min

72°C - 3.00 min

repeat 3-5 for 35 cycles

72°C - 10.00 min

4°C - forever


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