Part:BBa_K1085018:Experience

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Applications of BBa_K1085018

The part has been placed into the iGEM Groningen Bacillus subtilis integration backbone (BBa_K1085014) in order to test expression of the silk protein.
In order to prove that the following checks were made:
Starch assay was performed:

The upper 6 colonies are transformants of BBa_K1085018. The ones which don't show any halo confirm that the part integrated into the amyE locus, since amylase activity is disrupted: Bacillus subtilis is not able to process starch.

Colony PCR was also performed on the same 6 colonies using one primer which binds on the amyE locus and the other within the promoter region.
The samples were checked on agarose gel 0.8%.

The upper part of the picture shows the colonies which are supposed to contain the BBa_K1085018. The samples showed to be all positive candidates (expected product size ~2400bps).

User Reviews

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