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Team:Northwestern/Notebook/Week8
From 2011.igem.org
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Notebook
Week 8
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
