IGEM:IMPERIAL/2006/project/Oscillator/Problems/aiiA

The aiiA does not degrade AHL this is a major problem which must be solved. To me it seems that no-one else is actually trying to solve this problem so we need the whole groups input.

Data showing aiiA does not work


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 * Tom 13:55, 26 August 2006 (EDT): I'd be a little hesitant to condem AiiA with that data alone, for the following reasons:
 * 1) The control of having non-induced cells I don't think is adequate. Having non-induced cells as a control we are assuming the operator sequence is functioning and intact and we're also making the assumption there is enough native LacI repressor to effectively shut off AiiA production. We need a control where we have AHL innoculated with non-AiiA/LuxI/Flourescence producing cells.
 * 2) Secondly, the method for working out whether the difference between induced and non-induced cells is statistically significant is not given. Just because a difference is between 3-4% doesn't make it statistically insignificant by default. Especially if you're consistently getting those results. I'd suggest a T test on repeated samples to determine significance.
 * 3) Thirdly, the dilution of cells and small AHL innoculation time may be having an effect. I'd suggest growing a culture overnight in the prescence of AHL and IPTG, along with appropriate controls, and taking the supernatent directly from that for the T9002 assay.

Info about aiiA and new aiiA
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The new aiiA gene needs Primers which have Registary restriction sites and LVA tag, someone design them and put them here