Part:BBa_J100205:Experience

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Applications of BBa_J100205

J100205 (repClone Red) was used to build part BBa_J100306, repClone Red with wt tetR promoter. We transformed E. coli with plasmids containing J100306 or J100308 (a variant of J100306) and measured the induction caused by the addition of 200 ng/mL anhydrous tetracycline (aTc) to determine the incubation time needed for maximum induction. We found that aTc is most effective after fourteen to sixteen hours of growth, after which the induction is lost.

We performed an aTc dose-response test on E. coli containing part J100306 to determine the ideal concentration of aTc for induction of RFP. We found that 400 ng/mL aTc gives the best compromise of fold induction and allowing for sufficient cell growth. Induction above 400 ng/mL looks higher because fewer cells grew and therefore the denominator was smaller when calculating the ratio.

User Reviews

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