Competency refers to the ability of cells to take up free, extracellular genetic material (e.g., plasmid DNA). This article will tell you exactly how to achieve this and, hopefully, set you well on your way to yeast transformation success! The following protocol can be used to transform either freshly prepared or frozen competent Pichia cells. Transformation efficiency may vary with each strain and vector used. Biotechniques. 2005 Jan;38 (1):44, 46, 48. doi: 10.2144/05381BM04. 1. Add 10 μl DNA (5-10 μg) to 80 μl of electrocompetent cells. Gently mix with a blue-tip pipette several times. Transfer mixture into a prechilled cuvette and incubate on ice for 5 minutes. 2. Wipe moisture from the cuvette and insert the cuvette into the device. 3. Electroporation: 4. Immediately add 1 ml of ice-cold sorbitol to the cuvette. • It is also very important to keep the cells at 4°C for the remainder of the procedure. The cells, and any bottles or solutions that they come in contact with, must be pre-chilled to 4°C. The Pichia EasyComp™ Transformation Kit contains sufficient reagents (Solutions I, II, and III) for 6 preparations of competent cells. Each competent cell preparation yields enough cells for 20 transformations. doi: 10.1007/978-1-0716-2399-2_7 - Springer Table 1 compares the steps in cell preparation and transformation for conventional electroporation, heat shock, and our condensed protocol. Compared to the heat-shock method, the condensed protocol requires less time for the transformation step and provides much higher transformation efficiencies. We describe here a shortened protocol for cell preparation and transformation that works reliably with either prototrophic markers or antibiotic selection in this host. We describe here a shortened protocol for cell preparation and transformation that works reliably with either prototrophic markers or antibiotic selection in this host.