From 2009.igem.org
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| - | Descriptive Title of What You're Doing
| + | Visualization of Previous Restriction Digest |
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| - | WIKI CODING HERE
| + | *Visualization of psB1AC3, psB1AK3 and LuxOD47E in pCR.2.1-TOPO vector restriction digest |
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| | + | *Added 10X Orange dye and ran on a 1% agarose gel, expected band sizes: psB1AC3/psB1AK3 ~ 3.0 KB, |
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| | + | *Lane 1- Ladder |
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| | + | Lanes 2-6 psB1AC3 |
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| | + | Lanes 7-11 psB1AK3 |
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| | + | Lane 8- LuxOU |
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| | + | Lane 9-Lux PQ |
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| | + | Lane 10- Lux CDABE |
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| | + | Lane 11- LuxOD47E |
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| | + | Lane 12- LuxOD47A |
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| | + | *Results: Gel Photo |
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| | + | *Analysis: From this gel we can conclude that LuxPQ and LuxCDABE were not cut with EcoRI as we do not see the expected band sizes in these lanes. The others look good. We will proceed by sending LuxPQ in TOPO vector and LuxCDABE in TOPO vector for DNA sequencing. |
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Revision as of 02:54, 21 August 2009
University of Calgary
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CAROL
Descriptive Title of What You’re Doing
WIKI CODING HERE
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CHINMOYEE
More Simbiology
Exploration Results :
Simbiology assumes that all reactions are elementary reactions. For stochastic simulation only the Chemical kinetic and Hill kinetic equations work. Simbiology also produces a diagram of all the reactions input . The diagram view is helpfull in establishing the connections between species when they react or form.
The simbiology simulation graphs produced have a generic y-axis value of state which signifies concentration .
Started to look at other Toolboxes given with the Matlab package.
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EMILY
Visualization of Previous Restriction Digest
- Visualization of psB1AC3, psB1AK3 and LuxOD47E in pCR.2.1-TOPO vector restriction digest
- Added 10X Orange dye and ran on a 1% agarose gel, expected band sizes: psB1AC3/psB1AK3 ~ 3.0 KB,
Lanes 2-6 psB1AC3
Lanes 7-11 psB1AK3
Lane 8- LuxOU
Lane 9-Lux PQ
Lane 10- Lux CDABE
Lane 11- LuxOD47E
Lane 12- LuxOD47A
- Analysis: From this gel we can conclude that LuxPQ and LuxCDABE were not cut with EcoRI as we do not see the expected band sizes in these lanes. The others look good. We will proceed by sending LuxPQ in TOPO vector and LuxCDABE in TOPO vector for DNA sequencing.
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FAHD
Descriptive Title of What You're Doing
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IMAN
Descriptive Title of What You're Doing
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JAMIE
Descriptive Title of What You're Doing
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JEREMY
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KATIE
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KEVIN
Cloning Technique
Began looking into biobrick cloning techniques. Reviewed how the parts are put together. If A part is the recepient and B part is the donor, and when B part needs to be put behind of A, the recepient A is cut with SpeI and PstI sites and the donor is cut with XbaI and PstI. When B needs to be put in front of A, the recepient A is cut with EcoRI and Xba, and the donor B is cut with EcoRI and SpeI. When cut Xba and Spe sites meet, it creates a scar, making the construction permenant. (It won't cut apart with the enzymes we use)
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MANDY
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PATRICK
Descriptive Title of What You're Doing
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PRIMA
Descriptive Title of What You're Doing
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STEFAN
Descriptive Title of What You're Doing
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VICKI
Gel of the vector verification performed yesterday
Purpose: to visualise the results of yesterday’s vector verification
Expected restriction digest output:
Biobrick digest (2 fragments)
- psB1AC3/psB1AK3: ~3.0 kb
- ccdB (p1010): ~675 bp
TOPO digest (2 fragments)
- TOPO (T/A and Blunt): ~3.5 kb
- LuxOU: ~2.0 kb
- LuxPQ: ~4.0 kb
- LuxCDABE: ~6.0 kb
- LuxOD47E: ~1.4 kb
- LuxOD47A: ~1.4 kb
Results:
Gel and lane description is included below. LuxPQ did not work, so it will need to be sequenced.
Sequencing of LuxPQ in TOPO Blunt II
Purpose: As mentioned above, the results for LuxPQ are very questionable. Since they cannot be explained with restriction digests, hopefully sequencing will alleviate some of the confusion.
Materials and methods:
We inserted 7.3 uL of LuxPQ in TOPO into a mini PCR tube so that around 730 ng of DNA were present for sequencing. We used the sp6 forward primer and the T7 reverse primer.
Results:
Much like our previous sequencing attempts, these were questionable. The electronic copy will be inserted shortly.
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