Preparation and properties of insolubilized restriction endonucleases

Yan-Hwa Wu Lee*, Robert W. Blakesley, Leonard A. Smith, Jack G. Chirikjian

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

Type II restriction endonucleases Bam HI and Eco RI were covalently coupled to Sepharose. These insolubilized enzymes generated fragment patterns for several viral DNAs identical to those produced by the respective free enzymes. Conditions for optimal activity were similar for both bound and unbound forms of the enzymes. Insolubilization improved thermal stability of Bam HI and Eco RI. The bound enzyme can be recovered from reaction mixtures and reused several times. Upon storage at 4°C, coupled endonucleases remained stable for several months.

Original languageEnglish
Pages (from-to)679-689
Number of pages11
JournalNucleic acids research
Volume5
Issue number3
DOIs
StatePublished - 1 Mar 1978

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