Isolation of Restrictible DNA

Elizabeta Topić; Jelica Gluhak

doi:10.1515/cclm.1991.29.5.327

Isolation of Restrictible DNA

Elizabeta Topić, Jelica Gluhak

Research output: Contribution to journal › Article

22 Scopus citations

Abstract

A simple method for the Isolation of pure and highyield DNA from whole blood, suitable for restriction enzyme digestion, is described. The Steps of the procedure are as follows: Cell lysis with NH₄C1, 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 µg/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.

Original language	English (US)
Pages (from-to)	327-330
Number of pages	4
Journal	Clinical Chemistry and Laboratory Medicine
Volume	29
Issue number	5
DOIs	https://doi.org/10.1515/cclm.1991.29.5.327
State	Published - 1991
Externally published	Yes

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ASJC Scopus subject areas

Clinical Biochemistry
Biochemistry, medical

Cite this

Topić, E., & Gluhak, J. (1991). Isolation of Restrictible DNA. Clinical Chemistry and Laboratory Medicine, 29(5), 327-330. https://doi.org/10.1515/cclm.1991.29.5.327

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AB - A simple method for the Isolation of pure and highyield DNA from whole blood, suitable for restriction enzyme digestion, is described. The Steps of the procedure are as follows: Cell lysis with NH4C1, 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 µg/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.

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10.1515/cclm.1991.29.5.327