DNA extraction method for screening yeast clones by PCR.
A simple procedure for isolating yeast DNA suitable for use as a template for PCR amplification is described. SDS treatment alone is sufficient for extraction of chromosomal DNA from yeast cells. Cells of a yeast colony are suspended in a small volume (about 20 microL) of a 0.25% SDS solution, mixed vigorously and centrifuged. The supernatant can be directly used as a template after dilution to give an SDS concentration of less than 0.01% in the final PCR mixture.
Akada
R
R
Department of Applied Chemistry and Chemical Engineering, Faculty of Engineering, Yamaguchi University, Ube, Japan. rinji@po.cc.yamaguchi-u.ac.jp
Murakane
T
T
Nishizawa
Y
Y
D015825:Chromosomes, Fungal / Q000737:chemistry* / Q000235:genetics; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D004693:Glucan Endo-1,3-beta-D-Glucosidase / Q000737:chemistry; D007716:Kluyveromyces / Q000737:chemistry / Q000235:genetics; D017830:Octoxynol / Q000737:chemistry; D016133:Polymerase Chain Reaction / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000235:genetics; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry
D004271:DNA, Fungal; D012967:Sodium Dodecyl Sulfate; D017830:Octoxynol; D004693:Glucan Endo-1,3-beta-D-Glucosidase