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05/09/13
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#Heat shock the cells for 45 seconds in a 42'C water bath. Do not shake. Immediately return the tubes to ice for 2 minutes. | #Heat shock the cells for 45 seconds in a 42'C water bath. Do not shake. Immediately return the tubes to ice for 2 minutes. | ||
#Add 950 ul of SOC medium (at room temperature) to the ligation reactions. Incubate for 1.5 hours at 37'C on a shaker. | #Add 950 ul of SOC medium (at room temperature) to the ligation reactions. Incubate for 1.5 hours at 37'C on a shaker. | ||
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| + | **The controls for this experiment are the resistance and viability controls. One tube has 50 ul of highly competent cells and SOC medium added to it. | ||
Revision as of 11:10, 5 September 2013
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Transformation of pSB1C3/TOD genes
- Centrifuge the ligation
- Add 6 ul of each ligation reaction to a sterile 1.5ml tube on ice
- Thaw the cells on ice
- Carefully transfer 50ul of cells to the ligation reaction. Gently flick the tubes and incubate on ice for 20 minutes.
- Heat shock the cells for 45 seconds in a 42'C water bath. Do not shake. Immediately return the tubes to ice for 2 minutes.
- Add 950 ul of SOC medium (at room temperature) to the ligation reactions. Incubate for 1.5 hours at 37'C on a shaker.
- The controls for this experiment are the resistance and viability controls. One tube has 50 ul of highly competent cells and SOC medium added to it.
