Team:TU Delft/16 July 2010 content

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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:

# BioBrick Insert 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 25μL, 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. The following concentrations of plasmid were obtained:

BioBrick Concentration (ng/μL)
J23100 130.7
I13522 106.7