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A simple and efficient procedure for transformation of Schizosaccharomyces pombe
Author(s) -
Morita Tomotake,
Takegawa Kaoru
Publication year - 2004
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.1104
Subject(s) - transformation (genetics) , schizosaccharomyces pombe , tris , distilled water , polyethylene glycol , transformation efficiency , biology , dna , incubation , yield (engineering) , plasmid , chromatography , nuclear chemistry , microbiology and biotechnology , saccharomyces cerevisiae , materials science , biochemistry , yeast , chemistry , gene , agrobacterium , metallurgy
We describe a simple and efficient procedure for transformation of Schizosaccharomyces pombe. Sz. pombe colonies grown on minimal (SD) plates were directly removed and suspended in a 100 µl reaction mixture containing 70 µl PLATE solution (50% polyethylene glycol‐4000, 100 m M lithium acetate, 10 m M Tris–HCl, pH 4.9, and 1 m M EDTA), 10 µl plasmid DNA (1 µg), 10 µl carrier DNA (100 µg) and 10 µl sterile distilled water. After incubation at 30 °C for 1 h followed by heat shock treatment at 42 °C for 15 min, the reaction mixture was spread on a selection plate. The transformation efficiency obtained using the procedure was approximately 8000 transformants/µg DNA. The method is simple and time‐saving, making it especially useful for a large number of samples and when a high transformation efficiency is not required. Copyright © 2004 John Wiley & Sons, Ltd.