Isolation of Restrictible DNA

Elizabeta Topić, Jelica Gluhak

Research output: Contribution to journalArticle

21 Citations (Scopus)

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 languageEnglish (US)
Pages (from-to)327-330
Number of pages4
JournalClinical Chemistry and Laboratory Medicine
Volume29
Issue number5
DOIs
StatePublished - 1991
Externally publishedYes

Fingerprint

Digestion
DNA
Endopeptidase K
DNA Restriction Enzymes
Chloroform
Phenol
Edetic Acid
Clinical Chemistry
Blood
Ethanol
Genes
Enzymes
isopentyl alcohol

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

Isolation of Restrictible DNA. / Topić, Elizabeta; Gluhak, Jelica.

In: Clinical Chemistry and Laboratory Medicine, Vol. 29, No. 5, 1991, p. 327-330.

Research output: Contribution to journalArticle

Topić, Elizabeta ; Gluhak, Jelica. / Isolation of Restrictible DNA. In: Clinical Chemistry and Laboratory Medicine. 1991 ; Vol. 29, No. 5. pp. 327-330.
@article{48fbf5e2ce574033a7a6f7f8b63fa7da,
title = "Isolation of Restrictible DNA",
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.",
author = "Elizabeta Topić and Jelica Gluhak",
year = "1991",
doi = "10.1515/cclm.1991.29.5.327",
language = "English (US)",
volume = "29",
pages = "327--330",
journal = "Clinical Chemistry and Laboratory Medicine",
issn = "1434-6621",
publisher = "Walter de Gruyter GmbH & Co. KG",
number = "5",

}

TY - JOUR

T1 - Isolation of Restrictible DNA

AU - Topić, Elizabeta

AU - Gluhak, Jelica

PY - 1991

Y1 - 1991

N2 - 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.

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.

UR - http://www.scopus.com/inward/record.url?scp=84955845754&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84955845754&partnerID=8YFLogxK

U2 - 10.1515/cclm.1991.29.5.327

DO - 10.1515/cclm.1991.29.5.327

M3 - Article

AN - SCOPUS:84955845754

VL - 29

SP - 327

EP - 330

JO - Clinical Chemistry and Laboratory Medicine

JF - Clinical Chemistry and Laboratory Medicine

SN - 1434-6621

IS - 5

ER -