Protease K preparation without Ca2+, Whether the Protease K acitivity is affecte - help me! (Nov/24/2005 )

Now I am preparing the Protease K that is to be used in extracting Genomic DNA from human tissue. When I prepare the Protease K solution, according to the Molecular Cloning, A labratory manual,3rd edition, Calcium Acetate should be added, But I had no Calcium Acetate at hand, I wonder whether the Protease K activity is Affected without Ca2+ ions? whether the Calcium Chloride can act as the Calcium acetate substitute?

Any suggestion is appreciated? and I am very grateful to all of you!

You don't need Ca+ ions but ProtK will not be as active without. If you are doing a DNA extraction you don't need it- if you have EDTA in your buffer the Ca2+ will be chelated anyway so there is not point in adding it.

Daniel

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