Progress, Results, and Discussion
|Figure 1. R0 R50 Angiostatin Selection cycle course PCR. The gel shows that 10 cycles is the optimal number of cycles for this round of PCR. NTC shows some signs of primer amplification, but thankfully does not show an actual band.|
As stated above, the broken thermocycler was a big issue for this round. However, since the thermocycler has been fixed, hopefully another won't break right when I'm using it. The only other problem would be the fact that a single transcription kit will rarely have all the needed components, leading to an increase in the chance of a possible contaminate reaching the selection process.
Conclusion and Future Work
In the upcoming week, a PAGE gel will be run to determine if any RNA product was obtained. If a product was obtained, it will be quantified using nanodrop spectrophotometry. Results condensed in Table 1. Since it seems that filter based selection takes much less time than bead based selection, I am hopeful that I can get at least 1 round done per week. To prepare for assaying, I will start looking into radiation handling classes. If or when I obtain an aptamer by selection with Tris buffer, I will then obtain MES buffer and select for angiostatin under the conditions of the tumor microenvironment.
|Round||PCR Cycles||Peak Absorbance (nm)||Initial RNA Amount (pmol)||RNA Yield (pmol)|
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