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PMID: 7145714 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A simple and efficient method for the separation and detection of small DNA fragments by electrophoresis in formamide containing agarose gels and Southern blotting to DBM-paper.

Nucleic acids research ·Vol. 10 ·No. 19 ·1982-10-11 ·Pages 5753-63

Sun YL, Xu YZ, Chambon P

Abstract

We show in this report that DNA fragments smaller than 300 bp are separated with high resolution by electrophoresis in concentrated (up to 7%) agarose gels containing 50% formamide. The separated DNA fragments can subsequently be quantitatively transferred to DBM-paper [Alwine, J.C. et al., Proc. Natl. Acad. Sci. USA (1977) 74, 5350-54] using the Southern technique [Southern, E.M., J. Mol. Biol. (1975) 98, 503-517], while preserving the sharpness of the original gel pattern. Since thin (0.2-0.4 mm) and thick (up to 5 mm) agarose slab gels can be easily handled in vertical or horizontal apparatus, this method should prove to be a very useful extention of the Southern technique, applicable to a variety of analytical and preparative purposes.

MeSH Terms

Base Composition DNA/isolation & purification Diazonium Compounds Electrophoresis, Agar Gel/methods Formamides Indicators and Reagents Molecular Weight Nucleic Acid Hybridization Oligodeoxyribonucleotides/isolation & purification Oligonucleotides/isolation & purification Sepharose

Chemicals

Diazonium Compounds Formamides Indicators and Reagents Oligodeoxyribonucleotides Oligonucleotides diazobenzyloxymethyl groups formamide DNA Sepharose

Authors & Affiliations

3 authors, click to expand affiliations / ORCID

Sun Y L

Xu Y Z

Chambon P

References (11)

11 references, click to expand

Detection of specific sequences among DNA fragments separated by gel electrophoresis.
J Mol Biol. 1975 Nov 5;98(3):503-17 PMID: 1195397
Ovalbumin gene is split in chicken DNA.
Nature. 1977 Nov 24;270(5635):314-9 PMID: 593351
Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes.
Proc Natl Acad Sci U S A. 1977 Dec;74(12):5350-4 PMID: 414220
Transfer of small DNA fragments from polyacrylamide gels to diazobenzyloxymethyl-paper and detection by hybridization with DNA probes.
Biochem Biophys Res Commun. 1978 Dec 14;85(3):1104-12 PMID: 216362
Detection of specific RNAs or specific fragments of DNA by fractionation in gels and transfer to diazobenzyloxymethyl paper.
Methods Enzymol. 1979;68:220-42 PMID: 94421
Electrophoretic transfer of DNA, RNA and protein onto diazobenzyloxymethyl (DBM) - paper.
Nucleic Acids Res. 1980 Jan 25;8(2):299-317 PMID: 6158735
Electrophoretic transfer of proteins and nucleic acids from slab gels to diazobenzyloxymethyl cellulose or nitrocellulose sheets.
Anal Biochem. 1980 Mar 1;102(2):459-71 PMID: 7425302
Differential nuclease sensitivity of the ovalbumin and beta-globin chromatin regions in erythrocytes and oviduct cells of laying hen.
Nucleic Acids Res. 1980 Jun 25;8(12):2737-50 PMID: 6253891
Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose.
Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201-5 PMID: 6159641
Efficient transfer of highly resolved small DNA fragments from polyacrylamide gels to DBM paper.
Gene. 1980 Nov;11(3-4):283-90 PMID: 7215799
A reliable method for the recovery of DNA fragments from agarose and acrylamide gels.
Anal Biochem. 1981 Apr;112(2):295-8 PMID: 6266279

Article Info

Journal

Nucleic acids research

Abbr.

Nucleic Acids Res

ISSN

0305-1048

DOI

10.1093/nar/10.19.5753

Published

1982-10-11

Pages

5753-63

Language

English

Region

England

NLM ID

0411011

PMCID

PMC320927

Subset

External Links

PubMed source DOI: 10.1093/nar/10.19.5753 PMC Full Text View journal details

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A simple and efficient method for the separation and detection of small DNA fragments by electrophoresis in formamide containing agarose gels and Southern blotting to DBM-paper.

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