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Team:TU Delft/16 July 2010 content
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=Lab work= | =Lab work= | ||
| - | + | ==Ordered DNA + Solvent Tolerance and Hydrocarbon Sensing== | |
The colony PCR of [https://2010.igem.org/Team:TU_Delft#/blog?blog=15_July_2010 yesterday] was put on[[Team:TU_Delft/protocols/agarose_gel|1% agarose gel]] | The colony PCR of [https://2010.igem.org/Team:TU_Delft#/blog?blog=15_July_2010 yesterday] was put on[[Team:TU_Delft/protocols/agarose_gel|1% agarose gel]] | ||
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We used 3 mL of the bacterial cells to make [[Team:TU_Delft/protocols/freezing_bacterial_stocks|-80 °C stocks]]. | We used 3 mL of the bacterial cells to make [[Team:TU_Delft/protocols/freezing_bacterial_stocks|-80 °C stocks]]. | ||
| - | + | ==Characterization of Anderson RBS sequences== | |
| - | + | ====Fluorescence measurements Attempt #2==== | |
Data analysis to follow shortly (good stuff!) | Data analysis to follow shortly (good stuff!) | ||
| - | + | ====Assembly of reference construct & positive control==== | |
[https://2010.igem.org/Team:TU_Delft#/blog?blog=15_July_2010 Yesterday's] digestion products of K081005 and I13401 were set for ligation over the weekend at 4 °C: | [https://2010.igem.org/Team:TU_Delft#/blog?blog=15_July_2010 Yesterday's] digestion products of K081005 and I13401 were set for ligation over the weekend at 4 °C: | ||
Revision as of 19:31, 2 August 2010
Contents |
Lab work
Ordered DNA + Solvent Tolerance and Hydrocarbon Sensing
The colony PCR of yesterday was put on1% agarose gel
We harvested the 1 mL bacterial cells of possible correct transformations. The pellets were stored at -20 °C. We used 3 mL of the bacterial cells to make -80 °C stocks.
Characterization of Anderson RBS sequences
Fluorescence measurements Attempt #2
Data analysis to follow shortly (good stuff!)
Assembly of reference construct & positive control
Yesterday's digestion products of K081005 and I13401 were set for ligation over the weekend at 4 °C:
| # | BioBrick | Fragment | Recipient plasmid | Final volume |
| 1 | [http://partsregistry.org/wiki/index.php?title=Part:BBa_K136011 K136011] | 17.5 μL ‘E-K081005-S’ | 4 μL ‘E-pSB1A2 I13401-X’ | 25 μL |
| 2 | [http://partsregistry.org/wiki/index.php?title=Part:BBa_K136011 K136011] | 17.5 μL ‘E-K081005-S’ | 4 μL ‘E-pSB1A2 I13401-X | 25 μL |
| 3 | Ligation control | None | 4 μL ‘E-pSB1A2 I13401-X | 25 μL |
To all samples with an end volume of 25 μL, 2.5 μL Ligase buffer was added.
The Birnboim method was used to isolate J23100 and I13522 (both in pSB1A2) from the o/n 5 mL LB cultures. We used 3 mL of the bacterial cells to make -80 °C stocks . The following concentrations of plasmid were obtained:
| BioBrick | Concentration (ng/μL) |
| [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23100 J23100] | 130.7 |
| [http://partsregistry.org/wiki/index.php?title=Part:BBa_I13522 I13522] | 106.7 |
