840:153g:Projects/project5/2009/02/26

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A New Project

  • Because our E.P. gel yesterday showed us that we didn't have any of the parts required to continue with that project we've been forced to, yet again, redesign our project.
  • Our project is now based on a project from a previous semester. It's goal was to extract a gene that produced a blue/purple pigment and place in in E. Coli so that when the precursor in the pathway was placed in the media this gene would express a blue color in E. Coli.
  • There project ended with startling results. They found that the sequence that had extracted and sequenced, contained either an intron or a mutated area that yielded a restriction site. They discovered this when they ran a gel and found two parts where there should have been one.
  • We discussed two options that could further this project.
       1. We try to extract RNA from the orchid. (Because the intron, if it is indeed an intron,
would be removed from that product.
       2. Or we should extract DNA from the orchid, design primers to isolate only the area of
DNA that wasn't previous sequenced, check that area for interons, and then remove the interons or
restriction site using a method called "skip pcr".
  • We chose to take the second option because of the instability of RNA. We decided that we would rather do the DNA extraction because DNA is stable, much easier to work with, and we have a protocol for extracting it from that plant that we know works.
  • After we deliberated over our options and decided, we then made the buffers that we are going to need for the extraction we will do on Tuesday.
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