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From 2012.igem.org

Weeks:

June			July				August				September				October				November

Week 15: 17th September - 23rd September

this week we have worked to characterize Promotors and Reporters in order to test their validity by injection in tadpoles :

• biobricking promotors in Pcs2+ which are then to be characterized in tadpoles after cloning upstream to various reporters including mCFP, SfGFP, and citrine which are:
  • P Elastase
  • Hsp70
  • P Car A
  • FLk1
  • P CMV
  this work included standard PCR amplification, purification , plasmid digestion, ligation of vector-insert, transformation, miniprepping, and sequencing.

following that cloning the reporters downstream to various promotors was done

cloning og IAAh and IAAM in both Psb1C3 and Pcs2+ by standard biobrick portocol

Golden gate assembly of IAAM-IAAH cassette in both Psb1C3 and Pcs2+

Injection in tadpoles for the parts we managed to clone and was validated correctly by sequencing