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Antarctic Phosphatase is used to catalyze the removal of 5' phosphate from DNA. If I'm using it in the lab on a sample of 1-5 micrograms of digested DNA, how would I figure out how long to incubate the sample at 37 degrees Celsius?

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  • $\begingroup$ I typically just react it for an hour, that amount of DNA doesn't seem excessive. $\endgroup$
    – user137
    Mar 11, 2015 at 23:29
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    $\begingroup$ This set of protocols was really helpful for me when I got started cloning. $\endgroup$
    – user137
    Mar 12, 2015 at 0:15
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    $\begingroup$ VTC because not looking up the manufacturer protocol counts in my opinion as an unresearched homework question. $\endgroup$
    – March Ho
    Mar 12, 2015 at 7:38

1 Answer 1

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For 5' extensions or blunt ends, incubate the sample for 15 minutes at 37 degrees Celsius.

For 3' extensions, incubate the sample for an hour.

https://www.neb.com/protocols/1/01/01/vector-dephosphorylation-protocol

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  • $\begingroup$ Depends on amount of enzyme added. $\endgroup$
    – AliceD
    Mar 12, 2015 at 9:44

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