Team:Caltech/Week 7

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Revision as of 19:04, 28 July 2010

iGEM 2010

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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.

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.

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 ==Monday 7/26==
+[[Image:CIT-cpcr-gel1-7-26.jpg|200px|thumb|CPCR gel featuring L16, L19, L23, L24, L31 & L34. (7/26)]]
+[[Image:CIT-cpcr-gel2-7-26.jpg|200px|thumb|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!
@@ Line 26: / Line 28: @@
 * 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:
+* Ran 2 gels of today's CPCR products.
 ==Tuesday 7/27==
+[[Image:CIT-cpcr-gel-7-27.jpg|200px|thumb|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