Team:Newcastle/26 July 2010
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- | == | + | ==Preparation for cloning of the ''rocF'' BioBrick== |
- | ====Aim: | + | |
+ | ==Colony PCR of Genomic DNA== | ||
+ | |||
+ | ===Aim:=== | ||
To determine whether the genes have been inserted into the plasmid of ''B. Subtilis 168''. | To determine whether the genes have been inserted into the plasmid of ''B. Subtilis 168''. | ||
- | + | ===Materials:=== | |
* Pipette | * Pipette | ||
Line 18: | Line 21: | ||
* Forward and reverse primers | * Forward and reverse primers | ||
- | + | ===Protocol:=== | |
* For the full protocol, please refer to Colony PCR in Protocol List. | * For the full protocol, please refer to Colony PCR in Protocol List. | ||
- | + | ====Conditions in ThermoCycler:==== | |
* Melting temperature, Tm used for Anneal step is 59°C. | * Melting temperature, Tm used for Anneal step is 59°C. | ||
- | + | ===Results:=== | |
Gel electrophoresis will be run tomorrow to determine the results. | Gel electrophoresis will be run tomorrow to determine the results. | ||
- | + | ===Conclusion:=== | |
Please refer to Lab book dated 27.7.2010. | Please refer to Lab book dated 27.7.2010. | ||
{{Team:Newcastle/footer}} | {{Team:Newcastle/footer}} |
Revision as of 19:32, 26 July 2010
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Contents |
Preparation for cloning of the rocF BioBrick
Colony PCR of Genomic DNA
Aim:
To determine whether the genes have been inserted into the plasmid of B. Subtilis 168.
Materials:
- Pipette
- Microfuge
- Microtubes
- Distilled H2O
- Nucleotide DNTP
- 5x GoTaq buffer
- Template DNA (B. Subtilis ATCC 6633, 1:1 and 1:2)
- Forward and reverse primers
Protocol:
- For the full protocol, please refer to Colony PCR in Protocol List.
Conditions in ThermoCycler:
- Melting temperature, Tm used for Anneal step is 59°C.
Results:
Gel electrophoresis will be run tomorrow to determine the results.
Conclusion:
Please refer to Lab book dated 27.7.2010.
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