Team:Stockholm/28 June 2010
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= Andreas = | = Andreas = | ||
- | ==Colony PCR verification of pEX vector== | + | ===Colony gradient PCR verification of pEX vector=== |
- | + | Gradient PCR was run to optimize the colony PCR settings for designed pEX verification primers (pEXf and pEXr). Products analyzed by agarose gel electrophoresis. | |
- | === | + | ====Procedures==== |
- | == | + | |
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- | + | '''PCR settings''' | |
- | + | * Primers: | |
- | + | **pEXf forward primer (CGG CTC GTA TAA TGT GTG GAA TTG) | |
- | + | **pEXr reverse primer (CGT TCA CCG ACA AAC AAC AG) | |
- | + | * Gradient: 50°C, 55°C, 60°C | |
- | + | * Elongation time: 2 min 30 s | |
- | + | ** Expected amplicon size: 116 bp (pEXf) + 103 bp (pEXr) + 1188 bp (insert) = '''1407 bp''''' | |
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- | + | '''Agarose gel analysis''' | |
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*1% agarose gel | *1% agarose gel | ||
*2 ul sample | *2 ul sample | ||
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*170 V, 20 min | *170 V, 20 min | ||
- | + | ====Results==== | |
'''Figure to be added later.''' | '''Figure to be added later.''' | ||
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=Hassan= | =Hassan= | ||
May the "http://www.pathguide.org" guide us through the darkness :) | May the "http://www.pathguide.org" guide us through the darkness :) |
Revision as of 09:55, 6 July 2010
Contents |
Andreas
Colony gradient PCR verification of pEX vector
Gradient PCR was run to optimize the colony PCR settings for designed pEX verification primers (pEXf and pEXr). Products analyzed by agarose gel electrophoresis.
Procedures
PCR settings
- Primers:
- pEXf forward primer (CGG CTC GTA TAA TGT GTG GAA TTG)
- pEXr reverse primer (CGT TCA CCG ACA AAC AAC AG)
- Gradient: 50°C, 55°C, 60°C
- Elongation time: 2 min 30 s
- Expected amplicon size: 116 bp (pEXf) + 103 bp (pEXr) + 1188 bp (insert) = 1407 bp
Agarose gel analysis
- 1% agarose gel
- 2 ul sample
- 170 V, 20 min
Results
Figure to be added later.
Hassan
May the "http://www.pathguide.org" guide us through the darkness :)