Team:Northwestern/Notebook/Week8

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RETURN TO IGEM 2010


Day 34 - Monday, August 1st 2011


  • Miniprepped constitutive promoters, RBSs, and a 3 part ligation
  • Digested 2A (w/ PCR clean-up) and 3A for ligations of RBSs with parts
  • Ran gel of digested 3 part ligation
  • Transformed cells with ligated digestions
  • Found strep tag sequence? Fingers crossed. Worked on primers for adding tag to receptors.
  • Prepared samples for sequencing (lasP+RBS30+RFP, rhlP+RBS34+GFP)


Day 35 - Tuesday, August 2nd 2011


  • Made glycerol stock of constitutive promoter + RBS34 + rhlR
  • Digested, ligated, and transformed constitutive promoters and RBSs/parts
  • Finished primers for adding tag to receptors.
  • Prepared samples for sequencing.
  • PCRed a bunch of parts and 2L ligations; did ONs of backbones...we’re running out of DNA! =O
  • Received promoter oligos in mail!
  • Used up chlor plates for transformations so we made more plates (chlor and tet)
  • Plated ligations from glycerol stocks


Day 36 - Wednesday, August 3rd 2011


  • Miniprepped yesterday’s ONs
  • Made glycerol stock of CP2
  • Annealed CP oligos received in mail
  • Digested RBSs+parts and ligated with CP oligo
  • Most of yesterday’s transforms did not work - ONed transforms that did work


Day 37 - Thursday, August 4th 2011


  • Miniprepped, digested, and ran gels of RBS34+parts and RBS30+parts ligations
  • Miniprepped chlor backbone
  • Transformed oligo ligations
  • Plated RBS30+rhlR from glycerol stock
  • Plated TOP10 cells seedstock in preparation for making more competent cells


Day 38 - Friday, August 5th 2011


  • Made new sterile MgCl2
  • Made glycerol stocks of what worked on yesterday’s gel
  • Worked on our super awesome iGEM t-shirt design