From 2010.igem.org
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| | ==Tuesday 7/27== | | ==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. |
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| | ==Wednesday 7/28== | | ==Wednesday 7/28== |
Revision as of 18:22, 28 July 2010
iGEM 2010
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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
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