Rapid isolation of mouse DNA from cells in tissue culture
W Meinke, DA Goldstein, MR Hall - Analytical Biochemistry, 1974 - Elsevier
W Meinke, DA Goldstein, MR Hall
Analytical Biochemistry, 1974•ElsevierA rapid method for the isolation of DNA from mouse cells grown in tissue culture is
described. Cells are lysed in 0.24 m sodium phosphate, pH 6.8 buffer, containing 1%
sodium dodecyl sulfate, 8 m urea, and 10− 3m ethylene-diaminetetraacetic acid, and the
crude lysate applied to a column of hydroxyapatite. RNA and proteins are removed from the
column with 0.24 m sodium phosphate buffer containing 8 m urea while DNA is selectively
eluted with 0.48 m sodium phosphate buffer. There is almost total recovery of cellular DNA …
described. Cells are lysed in 0.24 m sodium phosphate, pH 6.8 buffer, containing 1%
sodium dodecyl sulfate, 8 m urea, and 10− 3m ethylene-diaminetetraacetic acid, and the
crude lysate applied to a column of hydroxyapatite. RNA and proteins are removed from the
column with 0.24 m sodium phosphate buffer containing 8 m urea while DNA is selectively
eluted with 0.48 m sodium phosphate buffer. There is almost total recovery of cellular DNA …
A rapid method for the isolation of DNA from mouse cells grown in tissue culture is described. Cells are lysed in 0.24 m sodium phosphate, pH 6.8 buffer, containing 1% sodium dodecyl sulfate, 8 m urea, and 10−3m ethylene-diaminetetraacetic acid, and the crude lysate applied to a column of hydroxyapatite. RNA and proteins are removed from the column with 0.24 m sodium phosphate buffer containing 8 m urea while DNA is selectively eluted with 0.48 m sodium phosphate buffer. There is almost total recovery of cellular DNA from the column and the DNA is virtually free from proteins and RNA. This extraction procedure indicates that a mouse cell contains about 8 × 10−6 μg of DNA.
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