Part:BBa_K2669002:Experience

Transformation results

Note that the results was gotten with the plasmid design represented in the composite part IDT optimised AmilCP sequence

Figure 1:Transformation plate of the IDT optimized AmilCP sequence. The circled colonies represent the colonies used in the stability assay.

Figure 2:Transformation plate of the original AmilCP sequence. The circled colonies represent the colonies used in the stability assay.

<p>Stability assay

Stability assay through liquid experiment was performed with all three plasmids in the IDT supplied backbone. 1 mL of LB with ampicillin (25 ug/ml) was inoculated with one single colony in eppendorf tubes in 10 replicates for each part. In order to allow 10 generations of growth, a 1:1000 dilution was made and incubated for 24 h. The colour comparison was done through visualization of centrifuged cultures and by analyzing the color intensity we could deduce the stability of the protein.

Figure 3: The result after 10 generations. The upper row represent 10 different colonies of the IDT optimized sequence of AmiLCP. The lower row represents the original AmilCP sequence.

Interpretation

Already after 10 generations it was clearly visible that more IDT optimized colonies had the strongest color compared to the original amilCP colonies. This indicates that the stability of the IDT optimized AmilCP is greater than for the original AmilCP sequence.

Generation	IDT Optimized AmilCP Colony 1	IDT Optimized AmilCP Colony 2	IDT Optimized AmilCP Colony 3	IDT Optimized AmilCP Colony 4	IDT Optimized AmilCP Colony 5	IDT Optimized AmilCP Colony 6	IDT Optimized AmilCP Colony 7	IDT Optimized AmilCP Colony 8	IDT Optimized AmilCP Colony 9	IDT Optimized AmilCP Colony 10
10	+++	++	+++	+++	++	++	+++	+++	+++	+

Table 1: Interpretation of color expression of the IDT optimised AmilCP sequence after 10 generations of growth. +++ represents strong color, ++ represents moderate color, + represents weak color and - represents no color.

Generation	Original AmilCP Colony 1	Original AmilCP Colony 2	Original AmilCP Colony 3	Original AmilCP Colony 4	Original AmilCP Colony 5	Original AmilCP Colony 6	Original AmilCP Colony 7	Original AmilCP Colony 8	Original AmilCP Colony 9	Original AmilCP Colony 10
10	+++	+++	+++	+++	+++	+++	+++	+++	+++	+++

Table 2: Interpretation of color expression of the original AmilCP sequence after 10 generations of growth. +++ represents strong color, ++ represents moderate color, + represents weak color and - represents no color.

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