I am facing this weird problem in Pichia transformation. I am trying to transform a construct with both pPICZ backbone and pGAPZ backbone. pPICZ construct as well as vector control is transformed perfectly with abundant clones but pGAPZC construct and pGAPZ backbone both are giving only one or two colonies.
Same batch of competent cells. same amount of DNA 4-5ug and incubating for 1 hr in sorbitol after electroporation and another 2 hrs after adding YPD. plating on YPDS plates with 100ugml zeocin concentration.
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