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Day 1, 23.9.2014 Download APE.exe and familiarize Look for Trypanosoma cruzi hexokinase in GenBank or in the TriTryp database

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slide1

Day 1, 23.9.2014

  • Download APE.exe and familiarize
  • Look for Trypanosomacruzihexokinase in GenBankor in theTriTryp database
  • synthesizeforwardand reverse oligonucleotides to amplifythewhole gene, insertingsuitablerestriction sites in the 5´and 3´ ends, start with BamH1 (5´) and Xho1 (3´) sites, but do not include thestopcodon.
slide2

Rest in peace

*16.9.1928 + 24.9.2014

  • Day 2, 25.9. 2014
  • Lookupthesequence for eGFPandsynthesizeoligoswith a 5´ Sal1 site and a 3´ Xho1 site
  • Include a 3x Ala ou Gly – linker in the 5´ oligo
  • Fuse in APE theamplifiedfragment to theTcHexokinasefromtheotherday