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Rai, Sudhir Kumar; Atwood-Moore, Angela; Levin, Henry L
Methods in molecular biology, 01/2018, Letnik: 1721Journal Article
The introduction of ectopic DNA, such as plasmids, into yeast cells has for decades been a critical protocol for the study of this eukaryotic model system. We describe here an efficient transformation procedure for use in the fission yeast Schizosaccharomyces pombe. This method relies on chemical agents (lithium acetate, and polyethylene glycol) and temperature stresses, which ultimately facilitate transfer of the genetic material through the cell wall and plasma membrane without significant impact on the transferred DNA or the recipient cell. Using this protocol, we consistently see transformation efficiencies between 1.0 × 10 and 1.0 × 10 transformants per microgram of the plasmid with 10 S. pombe cells. The principal benefits and advantages of this method are its simplicity, efficiency, and relative speed of completion.
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Leto | Faktor vpliva | Izdaja | Kategorija | Razvrstitev | ||||
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JCR | SNIP | JCR | SNIP | JCR | SNIP | JCR | SNIP |
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in: SICRIS
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