Improved control of partial DNA restriction enzyme digest in agarose using limiting concentrations of Mg super(++)

Partial digests of DNA for cloning experiments are usually performed using either the quantity of restriction enzyme of incubation time, or both, as controlling factors. However, the authors have found that more precise and reproducible partial digests are obtained when a limiting concentration of t...

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Bibliographic Details
Published in:Nucleic acids research 1989-01, Vol.17 (2), p.808-808
Main Authors: Albertsen, H M, Le Paslier, D, Abderrahim, H, Dausset, J, Cann, H, Cohen, D
Format: Article
Language:English
Subjects:
DNA
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Summary:Partial digests of DNA for cloning experiments are usually performed using either the quantity of restriction enzyme of incubation time, or both, as controlling factors. However, the authors have found that more precise and reproducible partial digests are obtained when a limiting concentration of the required cofactor Mg super(++) is used. This method is especially useful for partial digests of DNA contained in agarose. In agarose, diffusion time of the much smaller Mg super(++) ion is shorter than that of a restriction enzyme. The result is a more homogeneous digestion of the DNA throughout the agarose block.
ISSN:0305-1048