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Team:Groningen/Notebook/Wetwork 3August2012
From 2012.igem.org
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Primer list:<br> | Primer list:<br> | ||
1. Fnr promoter <br> | 1. Fnr promoter <br> | ||
| - | 2. | + | 2. LacI promoter <br> |
3. Lycopene <br> | 3. Lycopene <br> | ||
4. Violacein <br> | 4. Violacein <br> | ||
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- B. subtilis (hopefully) containing pSac-Cm+lycopene grew <br> | - B. subtilis (hopefully) containing pSac-Cm+lycopene grew <br> | ||
- all E. coli transformation showed quite amount of colonies. The E.coli containing pSB1C3+alsT promoter showed pink-white colonies, which white colony implies disruption on the iGEM Groningen 2010 gene, therefore we expected that our alsT promoter+GFP segment was inserted.<br> | - all E. coli transformation showed quite amount of colonies. The E.coli containing pSB1C3+alsT promoter showed pink-white colonies, which white colony implies disruption on the iGEM Groningen 2010 gene, therefore we expected that our alsT promoter+GFP segment was inserted.<br> | ||
| + | <br> | ||
| + | To check whether the B. subtilis contained pSac-Cm+lycopene, colony PCR was done by using Taq polymerase. Unfortunately no colony showed the expected fragment (picture not shown). <br> | ||
| + | <br> | ||
| + | PCR of Fnr promoter, lycopene, violacein, and LacI promoter was done using Phusion polymerase. Annealing temperature used = 58oC. No expected size fragment shown on the agarose gel. | ||
Revision as of 16:42, 7 August 2012
Elbrich: Re-do of transformation with BBa_R0011 from distribution kit 2012 and 2009.
Nisa
Received 6 primer-pairs for pigments and promoter candidates.
Primer list:
1. Fnr promoter
2. LacI promoter
3. Lycopene
4. Violacein
5. AmilCP
6. AmilGFP
Result from transformation :
- B. subtilis (hopefully) containing pSac-Cm+lycopene grew
- all E. coli transformation showed quite amount of colonies. The E.coli containing pSB1C3+alsT promoter showed pink-white colonies, which white colony implies disruption on the iGEM Groningen 2010 gene, therefore we expected that our alsT promoter+GFP segment was inserted.
To check whether the B. subtilis contained pSac-Cm+lycopene, colony PCR was done by using Taq polymerase. Unfortunately no colony showed the expected fragment (picture not shown).
PCR of Fnr promoter, lycopene, violacein, and LacI promoter was done using Phusion polymerase. Annealing temperature used = 58oC. No expected size fragment shown on the agarose gel.
