IGEM:Arizona State/2014/Notebook/asu igem 2014/2014/08/12

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8/12

  • Transformed ligation mixture
    1. Ligation mixture
    2. - Control
    3. + Control (ACC Plasmid)

PCR Amplification of ACC parts

First, the dry primers were made into 100 μM master stocks. 1 μL was added per .1 nm. For example, a 31.2 nm sample had 312 μL added. This master stock was then diluted into working stocks of 10 μM. This was done by adding 1 μL of master mix into 9 μL H2O. This was done for all eight primers , four forward and four reverse.


The ACC plasmid was then diluted from 54.2 μg/μL to about 1 μg/μL through a 1/50 dilution.


PCR mix for all four reactions (accA, accB, accC, and accD)


Ethanol

Got plasmid PACYC Duet-pdc-adhb in strain DH10b and strain BL21 from Nielsen's lab to use for ethanol production

  • Plated both in preparation to transform plasmid into BL21 Strain.


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