Rapid transfer of low copy-number episomal plasmids fromSaccharomyces cerevisiae to Escherichia coli by electroporation |
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Authors: | Laura Gunn Jac A Nickoloff |
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Institution: | (1) Department of Cancer Biology, Harvard School of Public Health, 665 Huntington Avenue, 02115 Boston, MA |
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Abstract: | A simple and reproducible method for transferring low copy-number episomal plasmids from yeast toEscherichia coli has been developed. Although slightly more time-consuming than direct transfer methods, which are effective with high copy
number plasmids, the method is significantly faster than methods that require purification of yeast DNA. Plasmid DNA is released
from yeast cells during brief treatments involving grinding with glass beads and heating. The treated yeast are cooled, electrocompetentE. coli is added, the mixture is electroporated, and transformants are selected using standard conditions forE. coli electrotransformation. The procedure typically yields sufficient transformants for most applications. |
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Keywords: | Yeast episomes plasmid rescue electrotransformation |
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