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Team:Warsaw/Calendar-Main/18 September 2009
From 2009.igem.org
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(New page: {{WarNotebook}} <!-- do not edit above me! --> <h3>Assembly of endosome detection module:</h3> <h4>Marcin</h4> Task 1: Transformation of TOP10 bacterial strain with [http://partsregistry....) |
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* Plasmids were isolated using the A&A plasmid mini kit. Detailed procedure of the isolation is described [http://aabiot.com/products/dna_purification/plasmid_dna/plasmid_mini/protocol_plasmid_mini.pdf here] | * Plasmids were isolated using the A&A plasmid mini kit. Detailed procedure of the isolation is described [http://aabiot.com/products/dna_purification/plasmid_dna/plasmid_mini/protocol_plasmid_mini.pdf here] | ||
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Methods: | Methods: | ||
* Reaction mixture composition: | * Reaction mixture composition: | ||
Latest revision as of 15:01, 26 September 2009
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Contents |
Assembly of endosome detection module:
Marcin
Task 1: Transformation of TOP10 bacterial strain with [http://partsregistry.org/Part:BBa_K177044BBa_K177044] on [http://partsregistry.org/Part:pSB1A3pSB1A3] plasmid Methods:
- Ligation mixture was thermally inactivated (done by Kuba)
- Detailed protocol of transformation is described here
Assembly of fusion proteins
Marcin
Task 1: Isolation of [http://partsregistry.org/Part:pSB1A3pSB1A3] plasmid containing [http://partsregistry.org/Part:BBa_K177028BBa_K177028] Methods:
- Plasmids were isolated using the A&A plasmid mini kit. Detailed procedure of the isolation is described [http://aabiot.com/products/dna_purification/plasmid_dna/plasmid_mini/protocol_plasmid_mini.pdf here]
Task 2: Restriction digest of previously isolated plasmids:
Methods:
- Reaction mixture composition:
3 μl purified plasmid DNA product 1 μl EcoRI (Fermentas) 1 μl PstI (Fermentas) 2 μl Buffer Tango (Fermentas) 13 μl MQ water
- Reactions were carried out 1.5 hour
Results: Really disappointing, none of the attemps to ligate the signal peptide were successful.
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