Hi, everyone!
I am trying to introduce some BstXI-linearised pPICZA-constructs into Pichia pastoris. I would like to introduce them by electroporation, as that is what I have used in the past. Nevertheless, the electroporation cuvettes in my new lab are different and people don't usually use this technique.
In the past, I have used BioRad's MicroPulser electroporator with 0.2cm cuvettes in Yeast setting (2000V); here, there are 0.4cm cuvettes and electroporator is an Eppendorf Electroporator 2510.
Anyone could help?
Thanks!
In the Eppendorf 2510 I've always used 1500 V to electroporate Pichia in 0.2 cm cuvettes. In theory, with 0.4 cm cuvettes you should double the volume of cells (to keep cross-sectional area constant) and double voltage to produce a similar voltage gradient across the electrodes, so I'd try upping V to 2500 (the maximum the instrument allows).
Hope it helps (and let me know if it works, as my 0.2 cm cuvette stock is depleting at an alarming rate but have plenty of unused 0.4 cm cuvettes :)
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