Team:Caltech/Week 7

Monday 7/26

• Started PCR reaction to extract the HSP (K112400) from its Berkeley Standard backbone (BBb). (Used same PCR parameters as CPCR from 7/22.
• Group meeting!
  • By next week: Need to design test-construct experiments, as they will be ready sometime early next week.
    • Can use PCR machine to achieve easy temperature gradients
    • Use spectrophotometer and eventually microscope to assay fluorescence.
    • Consider a plate assay?
  • AIBN issues:
    • Consider using low-concentration DMSO to help dissolve AIBN in LB/cell mixture.
    • Need to figure out how much unreacted (or otherwise) AIBN remains after mixing with cells.
    • Need to test crosslinking using hydrolyzed and epoxidized oil mixed with lysate and with cells to tweak concentration of AIBN needed, and to see what it makes.
  • TGase:
    • purchase urea?
    • To prevent a solid plug forming upon addition of TGase powder to sample, prepare liquid stock of concentrated TGase.
  • Wiki:
    • Should visit biofab.org for research material for human impact segment.
  • In general, we should set goals for both the end of SURF and for the end of the summer and determine what we need to do to get on track for them.
    • Printing in 2D
    • Working AND gate, at least in bulk fluid
    • Characterization of 3D hydrogel/plastic
    • Crosslinking data
• PCR purified K112400 PCR product & digested as plasmid insert.
• Transformed RFP bricks of verious resistances to use as backbone for future ligations
• Ran 2 gels of today's CPCR products:

Tuesday 7/27

• Began more CPCR on failed ones from yesterday: L16, L19, L31, L34, L35
• Began more digests of miniprepped DNA from K112400
• Made more electrocompetent cells. Began two test transformations.
• Transformed ligation products: L42, L46, L47 and the HSP.
• Began 11 new ligations:
  • L48: R0082 + I13504 + Cm
  • L31: L17 + K215000 + Kan
  • L34: L17 + L21 + Amp
  • L35: L18 + L21 + Amp
  • L22: K156012 + B0034 + Cm
  • L24: K156014 + B0015 + Cm
  • L20: C0077 + B0015 + Cm
  • L49: K112400 + L40 + Cm
  • L25: B0015 + M30109 + Tet
  • L26: R0077 + K124017 + Cm
  • L50: L18 + L27 + Amp
• Note: Need to upstream-digest more of: J23119, L17, K156014, B0015 for future ligations.

Wednesday 7/28

Thursday 7/29

Friday 7/30

Weekend 7/31-8/1