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| - | PCR | + | * Get the following excel: [[File:Team-EPF-Lausanne_Protocol_PCR.xls]] |
| + | * Fill in the pink fields with the appropriate values. | ||
| + | ** The annealing temperature is generally the lowest Tm. For long primers, use lowest Tm-5 | ||
| + | ** The extension time is 15-30s per 1kb | ||
| + | * Prepare the master mix by adding the elements in the following order: | ||
| + | ** H20 | ||
| + | ** 5x HF Buffer | ||
| + | ** dNTP | ||
| + | ** Polymerase (keep it cold!) | ||
| + | * Put 50ul of the master mix in a PCR tube for each different PCR | ||
| + | * Add the primers and the DNA | ||
| + | * Start the PCR! | ||
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