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Team:Warsaw/Calendar-Main/6 September 2009
From 2009.igem.org
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* Transformation of chemocompetent E. coli TOP10 strain | * Transformation of chemocompetent E. coli TOP10 strain | ||
Construct to transform: | Construct to transform: | ||
| - | * [http://partsregistry.org/Part:BBa_B0032<span style="color: black">BBa_B0032</span>] with [http://partsregistry.org/Part:BBa_E0022<span style="color: black">BBa_E0022</span>] connected to [http://partsregistry.org/Part:BBa_B0032<span style="color: black">BBa_B0032</span>] with [http://partsregistry.org/Part:BBa_C0051<span style="color: black">BBa_C0051</span>][http://partsregistry.org/Part:pSB1A3<span style="color: black">pSB1A3</span>] plasmid | + | * [http://partsregistry.org/Part:BBa_B0032<span style="color: black">BBa_B0032</span>] with [http://partsregistry.org/Part:BBa_E0022<span style="color: black">BBa_E0022</span>] connected to [http://partsregistry.org/Part:BBa_B0032<span style="color: black">BBa_B0032</span>] with [http://partsregistry.org/Part:BBa_C0051<span style="color: black">BBa_C0051</span>] on [http://partsregistry.org/Part:pSB1A3<span style="color: black">pSB1A3</span>] plasmid |
Methods: | Methods: | ||
* Detailed protocol of transformation is described [https://2009.igem.org/Team:Warsaw/Calendar-Main/5_July_2009 here] | * Detailed protocol of transformation is described [https://2009.igem.org/Team:Warsaw/Calendar-Main/5_July_2009 here] | ||
Revision as of 16:32, 8 September 2009
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Assembly of endosomal detection operon
Marcin
Task:1
- Transformation of chemocompetent E. coli TOP10 strain
Construct to transform:
- [http://partsregistry.org/Part:BBa_B0032BBa_B0032] with [http://partsregistry.org/Part:BBa_E0022BBa_E0022] connected to [http://partsregistry.org/Part:BBa_B0032BBa_B0032] with [http://partsregistry.org/Part:BBa_C0051BBa_C0051] on [http://partsregistry.org/Part:pSB1A3pSB1A3] plasmid
Methods:
- Detailed protocol of transformation is described here
Task 2:
- Digest isolated plasmid with Bax CDS to verify the correctness of the ligation:
Methods:
- Reaction mixture composition:
2 μl purified plasmid DNA product 1 μl RsaI (NEB) 2 μl Buffer Tango (Fermentas) 15 μl MQ water
- Digest was performed 3 hours
Results:
Comments:
- The enzym must have been deactivated due to presence of two forms of the plasmid. If circular plasmid is cut there are only one linear form in the sample. Althought in each case there was two forms - circular and supercoiled.
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