Team:Calgary/21 July 2010
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'''PCR Conditions''' | '''PCR Conditions''' | ||
*Cycle 1: 94°C 2:00 min | *Cycle 1: 94°C 2:00 min | ||
- | *Cycle 2: | + | *Cycle 2 * 36 cycles: |
**Stage 1:94°C for 1:00 min | **Stage 1:94°C for 1:00 min | ||
**Stage 2:52°C for 0:45 min | **Stage 2:52°C for 0:45 min |
Revision as of 22:03, 21 July 2010
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Wednesday July 21, 2010
Raida:
Objective: PCR out native MalE/ MalE 31 protein out of E.coli Purpose: Testing Mal E/ Mal E31 primers to get 1188 bp long gene of interest. Primers that are used are gene specific and anneals right on the start of the Mal E gene.
Tube Labels containing 1.0 μL of 10 mm DNA
- 1- p1H - C1
- 2- p1H - C1
- 3- p1H - C2
- 4- p1H - C2
- 5- p31H - C1
- 6- p31H - C2
- 7- p1ΔSS - C1
- 8- Master Mix
PCR Conditions
- Cycle 1: 94°C 2:00 min
- Cycle 2 * 36 cycles:
- Stage 1:94°C for 1:00 min
- Stage 2:52°C for 0:45 min
- Stage 3:72°C for 1:15 min
- Cycle 3: 72°C for 10:00 min
- Holding Temp: 4°C
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