840:153g:Projects/project2/2008/11/04

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The Sweet Smell of.....E.coli?

Today the ecoli group set the goal to do two things. the first one which i was involved with was to create another reverse primer for part number 45120. the primer that we made the first time did not make the part bio brick compatible. which requires two specific restriction sites to be in the front and back of the part. We made another reverse primer that will end up being 40 bp long. It will have the restriction site spe1 and pst1 on the end with the new primer.

The second goal was to try run a Gel that would be able to let us see the part 765001. The UV promoter part is only 76 bp long with around 15 to 20 bp on each end that seperate the part and the restriction sites. We cut the plasmid with two restriction enzymes. Ecor1 and Pst1 to see if a band of around 100 bp will show up on a 1.8% gel. We had tried it earlier with a 1% gel and we could not see the band that we wanted. We doubled the amount of DNA that we used for the restriction digest and for the Gel so that we would be able to see the band since it is small and were worried that we wouldn't be able to see it. We ran also to single digest of out plasmid to check if it would cut i the positions that we wanted. The result that we go showed a band just above the 100 marker line. We are really excited to see that result, because it tell us that we have the part at the expected size and can now concentrate on amplifying the part to use in out assembly of the bio brick part that will include a UV promoter and Wintergreen smell in a plasmid to put in to ecoli.