Revision as of 14:14, 20 June 2011

Linear template- TetR

Two step extension PCR

Put mix in PCR thermal cycler
Program the PCR thermal cycler: 1. 98°C for 30s 2. 98°C for 7.5 seconds 3. 58°C for 20 sec (this depends on the primers, therefore look up the annealing temperature of the primers used) 4. 72°C you have to calculate the time required. This polymerase has an effciency of 15-30s/kb. (calculate for the longest fragment) 6. 72°C for 5 min 7. 4°C "forever"
Save the file in the MAIN folder→ OK→ SAVE→RUN→Select the block you use→ RUN→ Change sample volume to 50�l, LidT should be 105°C and the box just below ("hotlid") should ALWAYS be checked
Press STATUS to check the progress

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 {{:Team:EPF-Lausanne/Templates/Header|title=Linear template- TetR}}
+Two step extension PCR
 == Materials ==
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 == Protocol ==
-* x.
+* Put mix in PCR thermal cycler
-* x
+* Program the PCR thermal cycler: <b>1.</b> 98°C for 30s <b>2.</b> 98°C for 7.5 seconds <b>3.</b> 58°C for 20 sec (this depends on the primers, therefore look up the annealing temperature of the primers used) <b>4.</b> 72°C you have to calculate the time required. This polymerase has an effciency of 15-30s/kb. (calculate for the longest fragment) <b>6.</b> 72°C for 5 min <b>7.</b> 4°C "forever"
+*Save the file in the MAIN folder→ OK→ SAVE→RUN→Select the block you use→ RUN→ Change sample volume to 50�l, LidT should be 105°C and the box just below ("hotlid") should ALWAYS be checked
+*Press STATUS to check the progress
 {{:Team:EPF-Lausanne/Templates/Footer}}