Team:Warsaw/Calendar-Stage2/21 September 2010

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Purification of MinC expression vectors

Kuba

Six colonies were picked for inoculation of liquid cultures (three with B0032 and three with B0034). Cultures were grown for 6 h in 37 ^oC. Plasmids were purified with Plasmid Mini kit (A&A) biotechnology. The obtained sample, as well as the initial (unmutated) plasmid, were then digested with EcoRI and PstI for 1 h and run on agarose gel.

M - size marker; 32a,b,c - plasmids harboring MinC under weak RBS; 34a,b,c - plasmids harboring MinC under strong RBS

Samples 32a,b&c ans well as 34b&c appear to contain a correct construct. Smaller amounts of DNA with B0034-containing constructs may indicate a slight leaking of the construct (probably due to a read-through from the resistance gene).