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PMID: 284385 Published · ppublish English Journal Article

Preparative and analytical purification of DNA from agarose.

Vogelstein B, Gillespie D

Abstract

Two procedures were developed for removing DNA from agarose after electrophoretic separation of DNA fragments according to size. Both involve dissolving the DNA-containing agarose in NaI. The preparative technique uses binding of DNA to glass in the presence of NaI. The method is rapid and convenient, and DNA of all molecular weight ranges can be recovered in high yield and without degradation. The DNA is free of agarose and remains susceptible to digestion by restriction enzymes. The analytical technique uses selective precipitation of DNA with acetone and has been adapted to molecular hybridization scans of sequences in agarose gels. The sequence-monitoring system is quantitative, directly measuring the proportion of the probe complementary to a given DNA fragment and vice versa. It is especially suitable for analyzing restriction enzyme digests of DNA in mapping experiments.

MeSH Terms
Acetone Base Sequence Chemical Precipitation DNA/isolation & purification DNA Restriction Enzymes/metabolism Electrophoresis/methods Electrophoresis, Agar Gel/methods Glass Humans Molecular Weight Sepharose/analysis Sodium Iodide Solubility
Chemicals
Acetone DNA Sepharose DNA Restriction Enzymes Sodium Iodide
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Vogelstein B
Gillespie D
References (18)
18 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
1979-02-00
Pages
615-9
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC382999
Subset
IM
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