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PCR
From 2011.igem.org
(Difference between revisions)
| Line 13: | Line 13: | ||
'''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 | ||
| + | |||
| + | 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 40 cycles | ||
| + | |||
| + | 72°C - 10.00 min | ||
| + | |||
| + | 4°C - forever | ||
[https://2011.igem.org/Team:Panama/Protocols/Wetlab '''Back'''] | [https://2011.igem.org/Team:Panama/Protocols/Wetlab '''Back'''] | ||
Revision as of 02:44, 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 40 cycles
72°C - 10.00 min
4°C - forever
