A simple, general procedure for purifying restriction endonucleases
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A simple, general procedure for purifying restriction endonucleases.
A simple, general method for purifying restriction endonucleases is described. The method employs precipitation of nucleic acids from crude extracts with polyethyleneimine followed by affinity chromatography on columns of heparin covalently linked to agarose. Most of the sixteen enzymes tested could be purified to a degree sufficient for DNA sequencing work by this method sometimes supplemented...
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Restriction endonucleases are highly specific in recognizing the particular DNA sequence they act on. However, their activity is affected by sequence context, enzyme concentration and buffer composition. Changes in these factors may lead to either ineffective cleavage at the cognate restriction site or relaxed specificity allowing cleavage of degenerate 'star' sites. Additionally, uncharacteriz...
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ژورنال
عنوان ژورنال: Nucleic Acids Research
سال: 1977
ISSN: 0305-1048,1362-4962
DOI: 10.1093/nar/4.8.2561