Transformation Success Tips

Transformation Success Tips - CaCl2 Method

Source plates must be fresh, between 16-19 hours old (try not to let plates go longer than 19 hours - refrigerate plates at 19 hours if necessary, do not use cells older than 24 hours, refrigerated or not).

Select enough cells (2-3 colonies @ 1 - 3 mm width) with toothpick or loop and make sure to avoid gauging agar. If past trials have failed, select more colonies.

Completely resuspend the cells in the CaCl2, leaving no cell clumps (vortex or mix vigorously to fully resuspend cells). Cell suspension should be slightly turbid or cloudy.

Prior to the heat shock at 42°C, extend incubation of cells in CaCl2 + DNA on ice (extra 10-15 min. - this allows more DNA to adhere to outside of cell).

Extend post-recovery at room temperature overnight, plate cells the next day - this extra recovery time will increase # of transformants.

If you have any other technical questions, please don't hesitate to contact us.

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