Part:BBa_K4683000:Experience

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Applications of BBa_K4683000

Lambert_GA 2022 Exponential Rolling Circle Amplification (eRCA) was successful with this part. The products of eRCA are short DNA strands composed of repeating complementary sequences of the used padlock probe. Therefore, one way in which the success of RCA can be determined is by running the exponential rolling circle products (eRCP) on an agarose gel. Since a fluorescent band very close to the wells would indicate the presence of an extremely long DNA strand, our RCP was run on a gel. The result was no/dim bands near the top of the well (see fig.1).

Figure 1. Image of gel ran with miRNA-1 RCP product.

Figure 1
By analyzing the results on the gel, our team concluded that short strands of DNA could be produced, likely the RCP, was produced.
The RCP was also tested with DFHBI-1T dye as the RCP would consist of Lettuce Aptamer sequences and the fluorescence was read on the plate reader (see fig.2).

Figure 2. Graph of split Lettuce reaction with RCP. The values represent the change in fluorescence before and after the reaction with DFHBI-1T took place.

As seen in Figure 2, the increase in fluorescence of the eRCP +dye was significantly greater than the controls, which suggests that the Lettuce produced. According to these results, eRCA was successful.

User Reviews

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