Projects Andy thought might yield results quickly.
1.) Identify different buffer conditions you might use for selection.
3.) Clone and sequence aptamers.
4.) Compare binding across different sets of conditions.
1.) Identify different buffer conditions you might use for selection.
- What are your variables? Ionic strength, pH, metal ions, temperature, what else? Which variables are most interesting or important to a successful selection?
3.) Clone and sequence aptamers.
4.) Compare binding across different sets of conditions.
- Do aptamers that bind at one ionic strength or pH still bind at another? How specific is evolution for a given condition? Are some variables more important than others?
- Has anyone done this before? What does the primary literature look like?
- Do the aptamers from a given pool or different pools bind at the same or different sites on lysozyme? How would you determine this? How might you do an ELISA? What is an ELISA?
- Do your experiments have any bearing on the origin of life? It is thought that modern life was preceded by a 'RNA world.' What is the RNA world hypothesis? Is a robust RNA world more likely under some environmental conditions than others? Did your experiments delimit the conditions under which RNA was more or less functional?
2 comments:
I did this! I looked at R0 background binding to lysozyme in 96 different buffers. I then selected 3 buffers that provided either high, normal (used the lysozyme buffer used in Colin Cox's previous selection), & low R0 background binding. I then performed 3 parallel anti-lysozyme aptamer selections. I can tell you more about this work, if you're interested.
-Gwen
Do you have any of the data? A paper? Post it up here and maybe someone can work off it. I remember you did the selection but did you test the binding of one aptamer in the other conditions? Ow specific was the aptamer to it's selection environment?
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