Composite

Part:BBa_K515102:Experience

Designed by: Atipat Patharagulpong   Group: iGEM11_Imperial_College_London   (2011-09-07)
Revision as of 17:12, 10 September 2011 by Nk2909 (Talk | contribs)

Characterisation

This part (BBa K515102) has been characterised qualitatively and quantitatively for chemotactic response towards L (-) malic acid. The tests were performed across a range of different attractant concentrations to observe optimal functioning of the part. Qualitative tests were performed using semi solid agar assays, quantitative analysis was performed using high - throughput capillary assay with cell count for each capillary evaluated using FACS. Behavioral analysis of E. coli expressing this part was done using Wide - field microscopy, and manual tracking was used to evaluate tactic response. Negative control used was E. coli 5α strain with a selectable marker for ampicillin and kanamycin. As positive control chemotactic response of E. coli 5α strain to attractant L (-) serine was tested, as serine is compatible with endogenous chemotaxis system of E. coli 5α strain.

Qualitative assay

Two tests were performed to observe chemotactic response over a large range of chemoattractant concentration and to observe differences in much finer milimolar ranges of attractant concentration.

Table 1: Ranges of attractant concentration tested.

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Attractant concentration

Test 1

0 M

10-5 M

10-4 M

10-3 M

10-2 M

10-1 M

Test 2

0 mM

5 mM

10 mM

15 mM

20 mM

25 mM


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