Isolation of restrictible DNA.
A simple method for the isolation of pure and high-yield DNA from whole blood, suitable for restriction enzyme digestion, is described. The steps of the procedure are as follows: cell lysis with NH4Cl, NaHCO3, EDTA; digestion with proteinase K in the presence of SDS; extraction with phenol-chloroform-isoamyl alcohol; and precipitation with ethanol. The 260 nm/280 nm absorbance ratio showed a mean value of 2, and the average yield of DNA was 212 micrograms/l. Such DNA preparations were found to be quite suitable for digestion by a variety of restriction endonucleases, as well as for the analysis of gene disorders by different biological methods. The method proposed appears to be useful in clinical chemistry laboratories.
Topić
E
E
Institute of Clinical Laboratory Diagnostics, Zagreb University School of Medicine, Yugoslavia.
Gluhak
J
J
D015139:Blotting, Southern; D004247:DNA / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D006801:Humans
D004247:DNA