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Team:Newcastle/15 July 2010
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==Materials== | ==Materials== | ||
| - | *[[Team:Newcastle/14_July_2010|Second round of | + | *[[Team:Newcastle/8_July_2010|Original transformants |
| + | *[[Team:Newcastle/14_July_2010|Second round of transformants]] | ||
*[[Team:Newcastle/12_July_2010|Plasmid extracts]] | *[[Team:Newcastle/12_July_2010|Plasmid extracts]] | ||
*Restriction enzymes ''Xba''1 aand ''Pst''1 | *Restriction enzymes ''Xba''1 aand ''Pst''1 | ||
==Protocol== | ==Protocol== | ||
| - | Single and Double | + | *Single (''Xba''1) and Double (''Xba''1 aand ''Pst''1) digest of Plasmid extracts |
| - | Run digests on a gel | + | *Run digests on a gel |
| - | Overnight culture of | + | *Overnight culture of all transformants |
==Results== | ==Results== | ||
| + | |||
| + | |||
| + | '''Figure 1.''' | ||
| + | |||
| + | This gel is unclear in precisely what is present in the samples it can clearly be seen that there are in fact two bands in the single digest lanes (where there should only be one) and three bands in the double digest lane (where there should only be two). | ||
| + | |||
| + | ==Conclusion== | ||
| + | |||
| + | It is our current belief that the [[Team:Newcastle/13_July_2010|plasmid extraction]] may have been performed incorrectly (there was one minor mistake during the procedure), so to this end an overnight culture of the original transformant colonies, as well as second round of transformants, will be performed, ready for plasmid extraction tomorrow. | ||
| + | |||
{{Team:Newcastle/footer}} | {{Team:Newcastle/footer}} | ||
Revision as of 14:52, 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
- [[Team:Newcastle/8_July_2010|Original transformants
- Second round of transformants
- Plasmid extracts
- Restriction enzymes Xba1 aand Pst1
Protocol
- Single (Xba1) and Double (Xba1 aand Pst1) digest of Plasmid extracts
- Run digests on a gel
- Overnight culture of all transformants
Results
Figure 1.
This gel is unclear in precisely what is present in the samples it can clearly be seen that there are in fact two bands in the single digest lanes (where there should only be one) and three bands in the double digest lane (where there should only be two).
Conclusion
It is our current belief that the plasmid extraction may have been performed incorrectly (there was one minor mistake during the procedure), so to this end an overnight culture of the original transformant colonies, as well as second round of transformants, will be performed, ready for plasmid extraction tomorrow.
   
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