From 2009.igem.org
Tuesday 7 July 2009
Results
Maria and Afton
- Overnight culture of I1352 grew, but no visible florescence
- Transformation Results below
| Name | Plate | Purpose | Growth Observation |
|
| NEB 10 beta
| LB | (+) Control shocked | lawn |
|
| NEB 10 beta
| LB | (+) Control | lawn |
|
| NEB 10 beta
| AMP | (-) Control | no growth |
|
| NEB 10 beta
| CM | (-) Control | no growth |
|
| NEB 10 beta
| CM | I1352 ligated to the backbone pSB4C5 | no growth |
|
| NEB 10 beta
| AMP | I1352 ligated to the backbone pSB4C5 | hundreds of colonies; some expressing RFP |
|
| NEB 10 beta
| AMP | I1352 ligated with itself | hundreds of colonies; some expressing RFP |
|
- Interpretation of Transformation Results:
- Shows the digestion/ligation process is not working
- Because the I1352 grew successfully on an AMP plate after being 'ligated' with pSB4C5, shows that the original backbone is present
- So it was not properly digested
- No growth was visible on the CM plate with the ligated I1352 with the CM resistant backbone pSB4C5
- If the digestion did not work properly, then the backbone could not be ligated with the I1352's promoter and RET
- Also, all previous stocks of BioBricks with promoters (100-107,110,113,116,0011,0040) have successfully grown on CM resistance plates, meaning they have the correct backbone, but none express RFP.
- Conclusion:
- The enzymes are bad for digestion
- The ligation process is a problem, because the original plasmid is going back into itself
- A new kit is being ordered
- Plans need to be made to repeat the Digestion and Ligation process with all promoters when kit arrives
Trentay 17:50, 31 July 2009 (UTC)
Tasks
Afton and Maria
Wetlab
Afton and Maria
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