TOP10 chemically competent cells: Difference between revisions

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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 yield a final OD of 0.25 in this test.  Typically this is around an additional 10 ml.
* Add chilled CCMB80 to yield a final OD of 1.0-1.5 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
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