Difference between revisions of "Part:BBa I13556:Experience"

Latest revision as of 21:26, 6 September 2007

Leakiness

It has been noticed that if this part is grown up in LB overnight in the absence of any inducer, the cells are strongly induced (up to about half the maximum fluorescence if AHL is added). This effect is not seen when the cells are grown up in EZ rich media or when the cells are made to grow slower. For example, there is a dramatic difference in the apparent leakiness of part depending on whether cells grown in LB are incubated at 37C (leaky) or 30C (not leaky). The root cause of this effect is unknown.

Transfer curve

Protocol

Two colonies of pSB4K5.I13556 were grown up in 5ml LB Kan overnight
Cultures were diluted 1:1000 and grown for 2 hours in 4ml EZ Rich medium + Kan
3 replicates of 100μl aliquots each for 8 AHL concentrations were made
added 100μl EZ+AHL to 8 final concentrations of AHL: 0, 1E-11, 1E-10, 1E-9, 1E-8, 1E-7, 1E-6, 1E-5 M
The cells were measured on a Victor3 fluorescence plate reader and grown overnight in the plate reader at 37C

Data

All data:

Taking the last time point for all samples, we get the following transfer curve (note that the first time point labeled as 1E-12 is actually 0 AHL and was just set there for ease of plotting):

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UNIQ8d40ea52ab6cea59-partinfo-00000000-QINU UNIQ8d40ea52ab6cea59-partinfo-00000001-QINU

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+==Leakiness==
+It has been noticed that if this part is grown up in LB overnight in the absence of any inducer, the cells are strongly induced (up to about half the maximum fluorescence if AHL is added). This effect is not seen when the cells are grown up in EZ rich media or when the cells are made to grow slower. For example, there is a dramatic difference in the apparent leakiness of part depending on whether cells grown in LB are incubated at 37C (leaky) or 30C (not leaky). The root cause of this effect is unknown.
 ==Transfer curve==