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Team:Newcastle/14 July 2010
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==Materials== | ==Materials== | ||
| - | *[[Team:Newcastle/13_July_2010| | + | *[[Team:Newcastle/13_July_2010|Overnight digests]] |
| - | + | ||
==Protocol== | ==Protocol== | ||
| - | + | The overnight digest are run on a gel | |
| + | Repeat overnight culture of colonies | ||
| + | Repeat transformation with ligation mix | ||
| + | |||
| + | ==Results== | ||
| + | The gel image was unfortunately lost, however the gel was run using all of the samples and a PCR product of known length, so as to check that the Molecular weight marker was running correctly. The results were the same as the gel run [[Team:Newcastle/13_July_2010|yesterday]], i.e. colonies 1 and 2 contain only pSB1AT3 vector (running at 3 Kbp) and the others contain pSB1AT3 with the RFP insert. | ||
| + | |||
| + | ==Conclusion== | ||
| + | The conclusion that can be reached from these results is that we have not been able to get transformants with the correct plasmid/insert combination. To this end the transformation is repeated. However there is again some confusion concerning the sizes, so the colonies undergo a second overnight culture, ready for a second plasmid extraction tomorrow. | ||
| + | |||
{{Team:Newcastle/footer}} | {{Team:Newcastle/footer}} | ||
Revision as of 11:59, 10 August 2010
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Contents |
LacI BioBrick Construction
Aims
- To use PCR to extract lacI (promoter, ribosome-binding site (RBS) & coding sequence (CDS)) from plasmid pMutin4 and ligate into vector pSB1AT3 in front of red fluorescent protein (RFP).
Materials
Protocol
The overnight digest are run on a gel Repeat overnight culture of colonies Repeat transformation with ligation mix
Results
The gel image was unfortunately lost, however the gel was run using all of the samples and a PCR product of known length, so as to check that the Molecular weight marker was running correctly. The results were the same as the gel run yesterday, i.e. colonies 1 and 2 contain only pSB1AT3 vector (running at 3 Kbp) and the others contain pSB1AT3 with the RFP insert.
Conclusion
The conclusion that can be reached from these results is that we have not been able to get transformants with the correct plasmid/insert combination. To this end the transformation is repeated. However there is again some confusion concerning the sizes, so the colonies undergo a second overnight culture, ready for a second plasmid extraction tomorrow.
   
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