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Team:ColegioFDR Peru/Solution

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Team:ColegioFDR Peru

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Our Solution/Methodology

 

The Keratinase sequence we decided to transform to E. coli (ATCC 25922) was a Ker(A or BPN) sequence found in Bacillus (Licheniformis or Methylotrophicus). We received the E. coli from an American Navy Research Hospital in Peru called NAMRU-6 (Naval Medical Research Unit Six) and we ordered the Keratinase sequence from the National Center for Biotechnology Information. Although we were never able to actually perform the experiment, the steps we were going to take in order to transform the E. coli involved: preparing the cells in order to make them “competent” or ready for transformation. Then, the actual protocol entails a series of procedures including freezing the cells at -70*C, followed by allowing the cells to thaw for 5 minutes, then incubating them for another 30 minutes on ice, followed heat shock at 37*C for 45 seconds, moving the cells back to ice for an additional 2 minutes, using a shaking incubator (225-250 rpm), and then finally incubating the cells at 37* C overnight. A more exact version of our hypothetically methodology can be found below:

 

 

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