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Team:TU Delft/16 July 2010 content
From 2010.igem.org
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(→Characterization of Anderson RBS sequences) |
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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]]. | ||
| - | + | <h4>Characterization of Anderson RBS sequences</h4> | |
| - | < | + | '''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: | + | |
| + | [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: | ||
{| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1" | {| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1" | ||
|'''#''' | |'''#''' | ||
|'''BioBrick''' | |'''BioBrick''' | ||
| - | |''' | + | |'''Fragment''' |
|'''Recipient plasmid''' | |'''Recipient plasmid''' | ||
|'''Final volume''' | |'''Final volume''' | ||
| Line 24: | Line 26: | ||
|[http://partsregistry.org/wiki/index.php?title=Part:BBa_K136011 K136011] | |[http://partsregistry.org/wiki/index.php?title=Part:BBa_K136011 K136011] | ||
|17.5 μL ‘E-K081005-S’ | |17.5 μL ‘E-K081005-S’ | ||
| - | |4 μL ‘E-pSB1A2 | + | |4 μL ‘E-pSB1A2 I13401-X’ |
|25 μL | |25 μL | ||
|- | |- | ||
| Line 30: | Line 32: | ||
|[http://partsregistry.org/wiki/index.php?title=Part:BBa_K136011 K136011] | |[http://partsregistry.org/wiki/index.php?title=Part:BBa_K136011 K136011] | ||
|17.5 μL ‘E-K081005-S’ | |17.5 μL ‘E-K081005-S’ | ||
| - | |4 μL ‘E-pSB1A2 | + | |4 μL ‘E-pSB1A2 I13401-X |
|25 μL | |25 μL | ||
|- | |- | ||
| Line 36: | Line 38: | ||
|Ligation control | |Ligation control | ||
|None | |None | ||
| - | |4 μL ‘E-pSB1A2 | + | |4 μL ‘E-pSB1A2 I13401-X |
|25 μL | |25 μL | ||
|} | |} | ||
| - | To all samples with an end volume of | + | To all samples with an end volume of 25 μL, 2.5 μL Ligase buffer was added. |
| - | The [[Team:TU_Delft/protocols/birnboim_plasmid_isolation|Birnboim method]] was used to isolate J23100 and I13522 (both in pSB1A2) from the o/n 5 mL LB cultures. The following concentrations of plasmid were obtained: | + | The [[Team:TU_Delft/protocols/birnboim_plasmid_isolation|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 [[Team:TU_Delft/protocols/freezing_bacterial_stocks|-80 °C stocks]] . The following concentrations of plasmid were obtained: |
{| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1" | {| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1" | ||
|'''BioBrick''' | |'''BioBrick''' | ||
Revision as of 17:55, 1 August 2010
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 | K136011 | 17.5 μL ‘E-K081005-S’ | 4 μL ‘E-pSB1A2 I13401-X’ | 25 μL |
| 2 | 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) |
| J23100 | 130.7 |
| I13522 | 106.7 |
