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Nucleic Acids Research, 1978, Vol. 5, No. 3 653-665
© 1978

Articles

Separation of very large DNA molecules by gel electrophoresis

Walton L. Fangman

Department of Genetics, SK-50, University of Washington Seattle, WA 98195, USA

Received December 16, 1977. Very large DNA molecules were separated by electrophoresis in horizontal slab gels of dilute agarose. Conditions of electrophoresis were developed using intact DNA molecules from the bacterial viruses {lambda}, T4 and G. Their DNAs have molecular weights (M) of 32 million, 120 million, and 500 million, respectively. Several electrophoresis conditions were found which give sufficiently high mobilities and large mobility differences that these DNAs are separated in a short time. Electrophoresis in 0.1% agarose at 2.5 V/cm of gel length separates T4 and {lambda} DNAs by 2.0 cm, and G and T4 DNAs by 1.0 cm in only 10 hr. With some conditions DNA mobilities are directly proportional to log M for M values from 10 to 500 million. The procedures. used will allow rapid molecular weight determination and separation of very large DNA molecules.


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