From 2009.igem.org
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| - | Descriptive Title of What You're Doing
| + | Plasmid isolation of overnight cultures and subsequent NotI restriction digest |
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| | + | Purpose: to confirm if a construct of the appropriate size can be excised from the plasmids transformed into TOP10 competent cells |
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| | + | Materials and methods |
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| | + | The plasmid isolation and restriction digest were performed in accordance with the procedure outlined on the protocol page. We expect to see 2 pieces from the J13002 + LuxOD47A BBk, one at 1685 bp and one at 3055 bp (for the AC plasmid); 2 pieces from the LuxOD47A BBk + B0015, one at 1750 bp and one at 3189 bp (for the AK plasmid); and 2 pieces from the LuxOD47A BBk size control, one at 1610 bp and one at 3055 bp (for the AC plasmid). |
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| | + | Results |
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| | + | Attached below. Lane 1 is the DNA ladder; lanes 2 and 3 are J13002 + LuxOD47A BBk (colonies 2 and 5 respectively); lane 4 is LuxOD47A BBk size control; lanes 5 and 6 are LuxOD47A BBk + B0015 (colonies 1 and 4 respectively); and lane 7 is a DNA ladder. The DNA ladder used was the GeneElute 1kb+. |
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Revision as of 23:18, 20 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
Final Exam
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EMILY
Descriptive Title of What You're Doing
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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
Descriptive Title of What You're Doing
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KATIE
Descriptive Title of What You're Doing
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KEVIN
Researching for a possible candidate of our response circuit
Mandy and I were assigned as partners and we were able to find a possible application of our signalling circuit involving the antifreeze proteins. There are variety of organisms that express Antifreeze proteins (AFPs), including fish (Arctic cod), bacteria (Pseudomonas putida), insects, and plants. Since we only work with bacteria, we attempted to find the most suitable antifreeze protein, and the antifreeze protein in Marinomonas primoryensis seemed to be most ideal.
Unlike other plant and bacterial antifreeze proteins, it is found to be hyperactive, meaning it is stable in low temperatures. Also, this antifreeze protein, like the one found in animals, utilizes a freeze-avoidant strategy which depresses the freezing point of the cell, while antifreeze proteins from other plants and bacteria utilize freeze-tolerant method, which does let the cell freeze, but it inhibits the growth of ice crystals; thus preventing the cell from damage.
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MANDY
Descriptive Title of What You're Doing
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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
Plasmid isolation of overnight cultures and subsequent NotI restriction digest
WIKI CODING HERE
Purpose: to confirm if a construct of the appropriate size can be excised from the plasmids transformed into TOP10 competent cells
Materials and methods
The plasmid isolation and restriction digest were performed in accordance with the procedure outlined on the protocol page. We expect to see 2 pieces from the J13002 + LuxOD47A BBk, one at 1685 bp and one at 3055 bp (for the AC plasmid); 2 pieces from the LuxOD47A BBk + B0015, one at 1750 bp and one at 3189 bp (for the AK plasmid); and 2 pieces from the LuxOD47A BBk size control, one at 1610 bp and one at 3055 bp (for the AC plasmid).
Results
Attached below. Lane 1 is the DNA ladder; lanes 2 and 3 are J13002 + LuxOD47A BBk (colonies 2 and 5 respectively); lane 4 is LuxOD47A BBk size control; lanes 5 and 6 are LuxOD47A BBk + B0015 (colonies 1 and 4 respectively); and lane 7 is a DNA ladder. The DNA ladder used was the GeneElute 1kb+.
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