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A colony procedure for transformation ofSaccharomyces cerevisiae

✍ Scribed by David Keszenman-Pereyra; Kotaro Hieda

Book ID
104741970
Publisher
Springer-Verlag
Year
1988
Tongue
English
Weight
261 KB
Volume
13
Category
Article
ISSN
0172-8083

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✦ Synopsis

A rapid and simple yeast transformation procedure has been developed using colonies on agar plates. Saccharomyces cerevisiae SHY3 cells were picked up from colonies on YPD plates grown freshly or stored at 4 degrees C and incubated with M13RK9-T DNA at 30 degrees C for 1-2 h in a solution of Li+, Ca2+, Mg2+, triacetin and polyethylene glycol. About 3,500 transformants were obtained per microgram of double stranded M13RK9-T DNA. Unlike the existing spheroplast techniques, single stranded M13RK9-T DNA transformed intact cells below one-hundredth frequency of the duplex form.

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