From 2009.igem.org
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- | The parts R0040, B0015, and J13002 were taken out from the 2009 DNA distribution kit, and were transformed into TOP10 cells. | + | The parts R0040, B0015, and J13002 were taken out from the 2009 DNA distribution kit, and were transformed into TOP10 cells. These parts are needed for general construction of our circuits. |
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Revision as of 22:19, 5 August 2009
University of Calgary
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CAROL
QuikChange XL Site-Directed Mutagenesis for ''luxCDABE''in TOPO vector
Purpose: To mutate the XbaI site that is located about ~1000bp into the sequence. The reason for mutating the XbaI site for luxCDABE in TOPO vector is because to clone the sequence into a biobrick vector (pSB1AK3 and pSB1AC3), the enzyme, 'XbaI' is required for cloning purposes. Instead of cutting the sequence into two parts, we have to mutate the site so that it only cuts at the end of the sequence properly.
- QuikChange XL Site-Directed Mutagenesis Kit retrived from Stratagene
- Both the control reaction and the control transformation was done as well to ensure efficiency of mutagenesis. Please see detailed procedures for mutagenesis in the protocol section.
- Plamids were transformed into XL Gold Ultracompetent cells (part of the kit) and was plated on LB plates overnight at 37oC. The XL Gold Ultracompetent cells allow the uptake of single stranded DNA.
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CHINMOYEE
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EMILY
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FAHD
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IMAN
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JAMIE
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JEREMY
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KATIE
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KEVIN
Bacterial transformation
The parts R0040, B0015, and J13002 were taken out from the 2009 DNA distribution kit, and were transformed into TOP10 cells. These parts are needed for general construction of our circuits.
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MANDY
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PATRICK
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PRIMA
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STEFAN
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VICKI
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