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Nucleic Acids Research, 1978, Vol. 5, No. 7 2223-2233
© 1978

Articles

Biospecific fractionation matrices for sequence specific endonucleases

Jay George and Jack G. Chirikjian

Department of Biochemistry, Georgetown University Medical Center, Georgetown University Washington, DC 20007, USA

Received May 22, 1978. Fractionation of several type II specific restriction endonucleases was achieved by separation on two novel biospecific matrices. The matrices are pyran, a copolymer of divinyl ether of maleic anhydride, and Cibacron Blue F3GA, a blue dye commonly used for the calibration of molecular sieves. Both compounds are insolubllized by coupling to sepharose through a cyanogen bromide linkage and in their soluble form inhibit the restriction endonucleases which we have tested. These affinity matrices can be used to obtain restriction endonucleases from crude extracts after removal of nucleic acids. They have also proven to have a high capacity when used as subsequent steps in enzyme purification. Their additional advantage is the rapid development time and reusability of columns packed with the two matrices.


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