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Team:Calgary/21 July 2010
From 2010.igem.org
(Difference between revisions)
Raidakhwaja (Talk | contribs) |
Raidakhwaja (Talk | contribs) |
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<u> Raida: </u> | <u> Raida: </u> | ||
| - | Objective: PCR out native | + | 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. | 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''' | + | '''Tube Labels containing 1.0 μL of 10 mm DNA''' |
*1- p1H - C1 | *1- p1H - C1 | ||
*2- p1H - C1 | *2- p1H - C1 | ||
| Line 18: | Line 18: | ||
'''PCR Conditions''' | '''PCR Conditions''' | ||
| - | Cycle 1: 94°C 2:00 min | + | *Cycle 1: 94°C 2:00 min |
| - | Cycle 2: | + | *Cycle 2: |
| - | *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 |
| - | *Stage 3:72°C for 1:15 min | + | **Stage 3:72°C for 1:15 min |
*Cycle 3: 72°C for 10:00 min | *Cycle 3: 72°C for 10:00 min | ||
| - | Holding Temp: 4°C | + | **Holding Temp: 4°C |
}} | }} | ||
Revision as of 22:02, 21 July 2010
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:
- 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
