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PCR
From 2011.igem.org
(Difference between revisions)
| Line 33: | Line 33: | ||
2. 94°C - 30 sec | 2. 94°C - 30 sec | ||
| - | 51°C - 1.00 min | + | 3. 51°C - 1.00 min |
| - | 72°C - 3.00 min | + | 4. 72°C - 3.00 min |
repeat 3-5 for 35 cycles | repeat 3-5 for 35 cycles | ||
| - | 72°C - 10.00 min | + | 6. 72°C - 10.00 min |
| - | 4°C - forever | + | 7. 4°C - forever |
[https://2011.igem.org/Team:Panama/Protocols/Wetlab '''Back'''] | [https://2011.igem.org/Team:Panama/Protocols/Wetlab '''Back'''] | ||
Revision as of 02:46, 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 3-5 for 35 cycles
6. 72°C - 10.00 min
7. 4°C - forever
