Team:LMU-Munich/Notebook/Protocols/24 Colony PCR
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(Difference between revisions)
(New page: {{:Team:LMU-Munich/Templates/Page Header}} Colony PCR {| |- |Component |Volume |Final concentration |- |PCR Master Mix, 2X |50 µl |1x |- |upstream primer | | |- |downstream primer ...) |
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|Component | |Component | ||
|Volume | |Volume | ||
- | |||
|- | |- | ||
- | |PCR Master Mix | + | |PCR Master Mix |
- | | | + | |2.5µl |
- | + | ||
- | |- | + | |- |
- | | | + | |primer forward (BioBrick) |
- | | | + | |0.375µl |
- | + | ||
|- | |- | ||
- | | | + | |primer revers (BioBrick) |
- | | | + | |0.375µl |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
|- | |- | ||
|nuclease free water to final volume of | |nuclease free water to final volume of | ||
- | | | + | |5µl |
- | + | ||
|} | |} | ||
+ | |||
+ | |||
+ | Guidelines for Amplification by PCR | ||
+ | |||
+ | :A. Denaturation: | ||
+ | ::initial: 94°C 5' | ||
+ | ::subsequent: 30'' | ||
+ | :B. Annealing: | ||
+ | ::53°C 30'' | ||
+ | :C. Extension | ||
+ | ::72°C 1'30'' | ||
+ | ::final: 8' | ||
+ | :D. Refrigeration | ||
+ | ::16°C | ||
+ | :E.Cycle Number | ||
+ | ::30 cycles | ||
+ | |||
+ | |||
+ | {{:Team:LMU-Munich/Templates/Page Footer}} |
Latest revision as of 12:47, 27 September 2010