pPICZαB-CalB

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	Plasmids were linearized within the AOX1 promoter with the restriction enzyme Mph1103I, purified with the DNA Clean & Concentrator™-5 (Zymo Research, Irvine, CA) after restriction digestion, and electroporated into competent yJC100 cells. Immediately after electroporation, cells were allowed to recover for 1 h at 30°C in 1ml of 50% 1M sorbitol/ 50% YPD and plated on selective media. Transformants were confirmed by colony PCR as described previously (Thor et al., 2005).

Protocol tips

Plasmids were linearized within the AOX1 promoter with the restriction enzyme Mph1103I, purified with the DNA Clean & Concentrator™-5 (Zymo Research, Irvine, CA) after restriction digestion, and electroporated into competent yJC100 cells. Immediately after electroporation, cells were allowed to recover for 1 h at 30°C in 1ml of 50% 1M sorbitol/ 50% YPD and plated on selective media. Transformants were confirmed by colony PCR as described previously (Thor et al., 2005).

Publication protocol

Yeast transformations were done by electroporation methods as described (Lin-Cereghino et al., 2005; Lin-Cereghino et al., 2007). Plasmids were linearized within the AOX1 promoter with the restriction enzyme Mph1103I, purified with the DNA Clean & Concentrator™-5 (Zymo Research, Irvine, CA) after restriction digestion, and electroporated into competent yJC100 cells. Immediately after electroporation, cells were allowed to recover for 1 h at 30°C in 1ml of 50% 1M sorbitol/ 50% YPD and plated on selective media. Transformants were confirmed by colony PCR as described previously (Thor et al., 2005).

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