Difference between revisions of "Part:BBa M36394:Experience"
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[[Image:secondary assay.jpg]] | [[Image:secondary assay.jpg]] | ||
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| + | These images show that from the experiments we conducted, 500uL is the ideal cell lysate volume to add to 500uL of Congo Red in order to maximize absorbance with 1000uM rhamnose. | ||
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| + | The Secondary Assay results do not demonstrate such a telling trend, but the data do seem to indicate that still 1000uM of rhamnose gives the highest absorbance after overnight growth. | ||
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| + | Please let us know if you attempt to isolate the protein or even spin it out!!! | ||
===User Reviews=== | ===User Reviews=== | ||
Revision as of 03:27, 7 December 2011
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Applications of BBa_M36394
Alhough we were unable to isolate spider silk protein produced by the E. coli cells, we did conduct dosage response assays on our system. We made use of Congo Red, a dye that binds to amyloid like proteins such as spider silk, to test for the presence of the protein in cell lysate at 541 nm. Altough we went off the methodology of Slotta et al. for usage of Congo Red, we were unable to isolate the spider silk protein in the limited time for the project. Thus, we conducted a primary assay to determine the ideal volume of cell lysate volume to add to Congo Red buffer before conducting a secondary assay adding this ideal volume of cell lysate to varying concentrations of rhamnose.
These images show that from the experiments we conducted, 500uL is the ideal cell lysate volume to add to 500uL of Congo Red in order to maximize absorbance with 1000uM rhamnose.
The Secondary Assay results do not demonstrate such a telling trend, but the data do seem to indicate that still 1000uM of rhamnose gives the highest absorbance after overnight growth.
Please let us know if you attempt to isolate the protein or even spin it out!!!
User Reviews
UNIQaa94ffe4ad626830-partinfo-00000000-QINU UNIQaa94ffe4ad626830-partinfo-00000001-QINU