User:Jonathan Dewerd/Notebook/TeamAllergy/2010/06/17

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Gel Extraction

Our intermediate-range goal is to express the hairpin that will suppress production of the isolated allergenic protein in E. Coli. The plan:

  1. Run a gel to isolate DNA strands of the desired length

Image:1kbLadderigem2010.JPG

  1. Perform a restriction digest of the PCR products and VO120 (our plasmid vector, Hi-Copy with Ampicilin) using Xba1 and Pst1.
  2. Ligate the result
  3. Transform it into E. Coli

Results

We ran a gel (see physical notebook for details) and then another gel, varying the temperature. Both indicated that we did not achieve the desired product: the allergenic gene was not being expressed in the strawberry. After further research, we hypothesize that this is because strawberries are non-climacteric -- that is, they reduce respiration after being picked (presumably reducing expression of the gene in question which is associated with coloration). We can try again with a fresher strawberry or perhaps change targets to a gene in tomatoes (which are climactic and exhibit a spike in respiration upon ripening).


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