User:Kchang17/notes: Difference between revisions
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**B0030.C2002.B0015.p8 (A) - innoculated at 0.0001 - grew ~14.5 hrs [[http://openwetware.org/wiki/Image:July22_2006_Reshmaverter_C2002.png picture]] | **B0030.C2002.B0015.p8 (A) - innoculated at 0.0001 - grew ~14.5 hrs [[http://openwetware.org/wiki/Image:July22_2006_Reshmaverter_C2002.png picture]] | ||
**B0030.C2003.B0015.p8 (B) - innoculated at 0.0001 - grew ~15 hrs [[http://openwetware.org/wiki/Image:July22_2006_Reshmaverter_C2003.png picture]] | **B0030.C2003.B0015.p8 (B) - innoculated at 0.0001 - grew ~15 hrs [[http://openwetware.org/wiki/Image:July22_2006_Reshmaverter_C2003.png picture]] | ||
*clones from July 6's penI red no induction culture, streaked on plate: | *clones from July 6's penI red no induction culture, streaked on plate: colony PCR of two red and two white colonies: | ||
[[Image:July8_2006_gel.jpg]] | |||
**insert should be ~1.6 kb | |||
**red colonies ~3 kb ([http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=149080 transposable element] jumped in) | |||
**white colonies ~1.6 kb | |||
**Q04740 red colony (C) - innoculated at 0.0001 - grew ~14.5 hrs [[http://openwetware.org/wiki/Image:July11_2006_penI_red.png picture]] | **Q04740 red colony (C) - innoculated at 0.0001 - grew ~14.5 hrs [[http://openwetware.org/wiki/Image:July11_2006_penI_red.png picture]] | ||
**Q04740 white (pink) colony (D) - innoculated at 0.0001 - grew ~14.5 hrs [[http://openwetware.org/wiki/Image:July11_2006_penI_white.png picture]] | **Q04740 white (pink) colony (D) - innoculated at 0.0001 - grew ~14.5 hrs [[http://openwetware.org/wiki/Image:July11_2006_penI_white.png picture]] | ||
*empty SP1.0 (E) - innoculated at 0.0001 - grew ~14.5 hrs [[http://openwetware.org/wiki/Image:July11_2006_emptySP1.png picture]] | *empty SP1.0 (E) - innoculated at 0.0001 - grew ~14.5 hrs [[http://openwetware.org/wiki/Image:July11_2006_emptySP1.png picture]] | ||
*Q04400.007 (F) - innoculated at 0.0001 - grew ~14.5 hrs [[http://openwetware.org/wiki/Image:July11_2006_tetR.png picture]] ([http://openwetware.org/wiki/Image:July11_2006_tetRconcat.png concatenated]) ([http://openwetware.org/wiki/Image:TetR_QPI.png MATLAB analysis]) | *Q04400.007 (F) - innoculated at 0.0001 - grew ~14.5 hrs [[http://openwetware.org/wiki/Image:July11_2006_tetR.png picture]] ([http://openwetware.org/wiki/Image:July11_2006_tetRconcat.png concatenated]) ([http://openwetware.org/wiki/Image:TetR_QPI.png MATLAB analysis]) |
Revision as of 15:39, 12 July 2006
February 28, 2006
- characterization of penI QPI in I13537.pSB1A3 with three replicates (replicate 1) (concatenated) (MATLAB analysis)
- rough measurement of original (unmutated) tetR inverter, Q04400 (one promoter SP1.0) (two promoter SP (I13537))
March 15, 2006
- first measurement of mutated tetR QPI Q04400.007 (obtained from inverter library) in SP1.0 (MATLAB analysis)
June 22, 2006
- tested everything in SP1.0 at 6 arabinose levels (0%-10-4%)
- Q04400.007 innoculated at OD 0.0005 and grew for ~10 hrs (picture) (MATLAB analysis)
- note: 10 hrs probably wasn't enough time for it to reach steady state esp. at high induction, assuming inverter was working better on July 11
- Q04740 exptl cultures grew too dense - performed an extra dilution and redid the innoculation - innoculated at OD 0.0005 and grew for ~14 hrs
- I14103 - grew very slowly - innoculated at OD 0.001 and grew for ~14 hrs (grew a little too dense)
June 29, 2006
- devices in SP1.0, six arabinose levels (0%-10-4%)
- Q04720 - innoculated at 0.0003 - grew ~15 hrs (too dense) (picture)
- Q04730 - innoculated at 0.0003 - grew ~16 hrs (too dense) (picture)
- during construction, a point mutation occurred in the ORF of the repressor protein (C0073) resulting in a change from Phe to Leu
- Q04740 - innoculated 0.0003 - grew ~16 hrs (too dense) - weirdness, where did GFP and RFP go?
- I14103 - innoculated at 0.0005 - grew ~15 hrs (too dense) (picture)
- I14104 - having trouble cloning this, have to wait on testing
July 6, 2006
- picture
- induced ~7 hrs at no and high induction
- the single red colony on a plate (probably got mixed with some white colony) from Q04740 in SP1.0 glycerol - got high RFP signal and GFP tracked induction - streaked this culture for July 11 testing
- one white colony from the same plate - GFP showed this time - background RFP
- empty SP1.0 - showed GFP and RFP
July 11, 2006
- grew to stationary, then diluted back 100x, grew for ~8 hrs (cultures were all very light except for the empty SP and Q04400.007), innoculated at ~11:30 p.m.
- devices in SP1.0, six arabinose levels (0%-10-4%)
- Reshma's constructs:
- clones from July 6's penI red no induction culture, streaked on plate: colony PCR of two red and two white colonies:
- insert should be ~1.6 kb
- red colonies ~3 kb (transposable element jumped in)
- white colonies ~1.6 kb
- Q04740 red colony (C) - innoculated at 0.0001 - grew ~14.5 hrs [picture]
- Q04740 white (pink) colony (D) - innoculated at 0.0001 - grew ~14.5 hrs [picture]
- empty SP1.0 (E) - innoculated at 0.0001 - grew ~14.5 hrs [picture]
- Q04400.007 (F) - innoculated at 0.0001 - grew ~14.5 hrs [picture] (concatenated) (MATLAB analysis)