IGEM:IMPERIAL/2009/Assays Protocols/Cloning Strategy M1&T

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Module 1 Cloning Strategy

II09 clonstratm1.jpg

  • Ligate PAH/ Cellulase gene sequence (G1) from GeneArt with the arabinose-inducible promoter from the iGEM starter kit.
  • Test for successful ligation using Cellulose/ PAH assay as defined here
  • If successful, remove terminators from G1 gene sequence using suitable restriction enzymes.
  • Ligate sequence to LacI (BBa_C0012) from the iGEM starter kit.
  • Test for expression of LacI gene using LacI repressible sequence coupled with reporter gene. BBa_I13601 is such a reporter - coupled with CFP.

II09 BBa I13601.jpg

Preliminary Schematic

II09 Clonstratm1 draft1.jpg