Transformations

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Step 2: Add plasmids and let the mixture sit in ice for 10 minutes
Step 2: Add plasmids and let the mixture sit in ice for 10 minutes
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Step 3: Quickly place in 42C water bath for 45 seconds
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Step 3: Quickly place in 42.C water bath for 45 seconds
Step 4: Return cells to ice for 2 minutes
Step 4: Return cells to ice for 2 minutes

Latest revision as of 15:59, 12 June 2006

This is the procedure we followed for transformation of competent E. coli cells.

Step 1: Thaw cells on ice

Step 2: Add plasmids and let the mixture sit in ice for 10 minutes

Step 3: Quickly place in 42.C water bath for 45 seconds

Step 4: Return cells to ice for 2 minutes

Step 5: Add 900 μl SOC and spin down, decant mixture

Step 6: Pipette 100 μl SOC and then plate out 90 μl

Step 7: Add 90 μl SOC to remaining 10 μl mixture and plate

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