High-throughput rapid yeast DNA extraction. Application to yeast artificial chromosomes as polymerase chain reaction templates.
An automation-assisted method enables DNA extraction of over a thousand yeast colonies in a day by one person. Yeast DNA is extracted essentially quantitatively in three steps: (a) cells are converted into spheroplasts with yeast lytic enzyme, (b) cells are lysed with proteinase K, and (c) DNA is collected by ethanol precipitation. The DNA stored at 4 degrees C is active in polymerase chain reaction experiments for more than a year.
Blanchard
M M
MM
Washington University School of Medicine, Center for Genetics in Medicine, St. Louis, Missouri 63110.
Nowotny
V
V
D018244:Chromosomes, Artificial, Yeast; D004271:DNA, Fungal / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction
D004271:DNA, Fungal