Transformation Protocol for ProFlex PCR

Thermocyclers makes transformations a lot easier by eliminating heat shocking the bacteria in a warm water bath.

I was hesitant to use the ProFlex PCR machine for transformations because its minimum temperature is 0°C. Previous transformations I’ve done via  thermocyclers always started the reaction at -4°C. I wasn’t sure if the 0°C would suffice. The manual protocol just says keep the samples on ice until ready for heat shock. I was searched the web for a transformation protocol for thermocyclers, but didn’t find anything substantial.

So I just did transformations with the ProFlex PCR machine to see what happens.

Turns out 0°C totally works! I’ve now done numerous transformations via the ProFlec PCR machine and they’ve worked. Having the lid heated vs unheated didn’t seem to make too much of a difference. I eventually opted for all unheated lid (just in case ambient heat was affecting my cells). I don’t have any data on its transformation efficiency versus a traditional method or versus a -4°C capable thermocycler. All I know is it worked for me.

Here’s the protocol:

5 min at 0°C #ramp down to 0°C, then load in the samples
30 min at 0°C
30 sec at 45°C
5 min at 0°C

Try it out! Let me know!

Don’t forget to put the PCR tubes in a cold block while pipetting!

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