Team:Bielefeld-Germany/Test

Revision as of 09:57, 26 October 2012 by Juvoss (Talk | contribs)

Week 26 (10/22 - 10/28/12)

Team Cellulose Binding Domain:
- Sequencing of <partinfo>BBa_K863120</partinfo> in <partinfo>BBa_J61101</partinfo> showed a deletion within the ORF of the GFP; this seemed to be the reason that the colonies are not fluorescent.
- Gradient-PCR on <partinfo>BBa_I13522</partinfo> with GFP_Freiburg Pre- and Suffix Primers
  - Worked fine. Clean up and digestion with XbaI and PstI
- Gradient-PCR on <partinfo>BBa_K863103</partinfo> with CBDcex_Freiburg and GFP_Freiburg_compl Primers
  - One of 12 temperatures worked. Clean up and digestion with XbaI and PstI
- Gradient-PCR on <partinfo>BBa_K863113</partinfo> with CBDclos_Freiburg and GFP_Freiburg_compl Primers
  - did not get product at all
- Ligation of the PCR products and GFP_Freiburg and CBDcex_Freiburg-GFP_Freiburg, respectively
- Transformation of both ligations
- Cells plated on selection agar

Team Site Directed Mutagenesis:
- Stopped Digestion of the shuttle vector, but gel went wrong and the products were lost

Team Cellulose Binding Domain:
- Colony PCR of B0034 + GFP_Freiburg
  - 14/24 positive
  - Plated three on selection agar for plasmid isolation
- Colony PCR of B0034 + CBDcex-GFP_Freiburg
  - 16/24 positive
  - Plated three on selection agar for plasmid isolation

Team Cellulose Binding Domain:
- Isolated Plasmids containing B0034 + GFP_Freiburg &
  - Digestion of pooled plasmids with XbaI and PstI
- Isolated Plasmids containing B0034 + CBDcex-GFP_Freiburg
  - Digestion of pooled plasmids with XbaI and PstI
- Digestion of J23100 with SpeI and PstI
- Clean up of inserts (B0034 + GFP_Freiburg & B0034 + CBDcex-GFP_Freiburg) and Backbone (J23100) via gel
- Ligation of Backbone and the inserts
- Transformation and plted on selection agar
Team Site Directed Mutagenesis:
- Digestion showed that all isolated colonies still have the illegal XbaI restriction side

Team Substrate Anaylsis: The MS-MS data arrived from Marcus Persicke. To ensure our measurements and discuss what happens after the Laccase treatment and on the ESI we decided to ask an organic chemistrer so we went to Prof. Dr. Dietmar Kuck whos expert on this field. Since he had less time to discuss we have only speculativ testifies.