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TOP10 chemically competent cells: Difference between revisions
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→Preparing competent cells
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* Centrifuge again at 4C and resuspend in 10 ml of ice cold CCMB80 buffer. | * Centrifuge again at 4C and resuspend in 10 ml of ice cold CCMB80 buffer. | ||
* Test OD of a mixture of 200 μl SOC and 50 μl of the resuspended cells. | * Test OD of a mixture of 200 μl SOC and 50 μl of the resuspended cells. | ||
* Add chilled CCMB80 to a final OD of 0.25 | * Add chilled CCMB80 to yield a final OD of 0.25 in this test. | ||
* Incubate on ice for 20 minutes | * Incubate on ice for 20 minutes | ||
* Aliquot to chilled screw top 2 ml vials or 50 μl into chilled microtiter plates | * Aliquot to chilled screw top 2 ml vials or 50 μl into chilled microtiter plates | ||
