From 2007.igem.org
yesterday -- tomorrow
PCR rxn
the following PCR reactions have been performed
| PCR : PrefixBB-frt1-CmR-frt2-SufixBB
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| PCR Settings
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| Buffer (5x)
| 5x 10µL (GC rich)
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| Expected size
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| Annealing (°C)
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| MgCl2 10µM
| 10µM 0µL (no extra)
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| 1Kb
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| 0min
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| dNTP 10µM
| 10µM 1µL
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| Success
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| Time Elongation
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| Oligo F 10µM
| O51
| 10µM 4µL
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| YES
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| 1m40'
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| Oligo R 10µM
| O52
| 10µM 4µL
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| Image (click to enlarge)
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| Number cycles
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| Water
| qsp 50µL
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| [[Image:|30px]]
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| 30
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| Polymerase
| Phusion 0,5µL
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| Band (0=ladder)
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| DNA
| pY-2P-IntC 5µL
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| PCR : PrefixBB-frt1-CmR-frt2-T1T2-SufixBB
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| PCR Settings
|
| Buffer (5x)
| 5x 10µL (GC rich)
|
| Expected size
|
| Annealing (°C)
|
| MgCl2 10µM
| 10µM 0µL (no extra)
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|
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| 0min
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| dNTP 10µM
| 10µM 1µL
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| Success
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| Time Elongation
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| Oligo F 10µM
| O51
| 10µM 4µL
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| NO
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| 1m40'
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| Oligo R 10µM
| O53
| 10µM 4µL
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| Image (click to enlarge)
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| Number cycles
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| Water
| qsp 50µL
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| [[Image:|30px]]
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| 30
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| Polymerase
| Phusion 0,5µL
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| Band (0=ladder)
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| DNA
| pY-2P-IntC 1µL (dil. 1/10)
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| PCR : PrefixBB-mutDGAT-SufixBB
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| PCR Settings
|
| Buffer (5x)
| 5x 10µL (GC rich)
|
| Expected size
|
| Annealing (°C)
|
| MgCl2 10µM
| 10µM 0µL (no extra)
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|
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| 0min
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| dNTP 10µM
| 10µM 1µL
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| Success
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| Time Elongation
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| Oligo F 10µM
| O29
| 10µM 4µL
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| NO
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| 2m
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| Oligo R 10µM
| O30
| 10µM 4µL
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| Image (click to enlarge)
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| Number cycles
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| Water
| qsp 50µL
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| [[Image:|30px]]
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| 33
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| Polymerase
| Phusion 0,5µL
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| Band (0=ladder)
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| DNA
| P13 1µL (dil. 1/10)
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