Team:Caltech/Week 7

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Monday 7/26

CPCR gel featuring L16, L19, L23, L24, L31 & L34. (7/26)
CPCR gel featuring L34, L35, L40, L42, L43 & L44. (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

CPCR gel featuring L16, L19, L31, L34, & L35. (7/22)
  • 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

  • Began CPCR reactions on the HSP, L42, L46, L48.

Thursday 7/29

Friday 7/30

Weekend 7/31-8/1

 
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