I'm curious: most protocols I see on transforming Pichia pastoris (which integrates a linear vector), calls for cleanup after linearization.
Basically, this is how the protocol goes:
- Linearize vector which contains gene of interest
- Cleanup linearization reaction, which apparently removes the salt and the restriction endonuclease used in the linearization process
- Combine the cleaned-up, linearized vector w/ electrocompetent yeast, and transform
- The linearized vector will be integrated into the genome, and this is how expression happens.
My question is: can I eliminate step 2? i.e. will leaving the salt and the enzyme in the transformation reaction ruin the transformation?