PCR

From 2011.igem.org

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'''Procedure:'''
'''Procedure:'''
-
1.
+
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
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 +
1. 94°C - 5.00 min
 +
 
 +
2. 94°C - 30 sec
 +
 
 +
3. 51°C -  1.00 min
 +
 
 +
4. 72°C - 3.00 min
 +
 
 +
repeat 2-4 for 35 cycles
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 +
6. 72°C - 10.00 min
 +
 +
7. 4°C - forever
[https://2011.igem.org/Team:Panama/Protocols/Wetlab '''Back''']
[https://2011.igem.org/Team:Panama/Protocols/Wetlab '''Back''']

Latest revision as of 02:48, 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

3. 51°C - 1.00 min

4. 72°C - 3.00 min

repeat 2-4 for 35 cycles

6. 72°C - 10.00 min

7. 4°C - forever


Back

Retrieved from "http://2011.igem.org/PCR"