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PacI digestion - (Aug/30/2006 )

Hi!!!!, somebody know if i can leave a digestion with PacI overnight... without problem of DNA degradation or star effect... or something bad....


it doesn't appear to be a problem, check this link

it does offer suggestions to increase efficiency under some conditions; you may want to follow the links if your digestion gave you any trouble

you can always go to and find out whether an enzyme exhibits star activity, and other properties


it is okay. Just make sure the DNA is clean, and the digestion carried out in sufficiently large volume, 100ul or more. (I am assuming you are cutting lots of DNA, thus requiring an overnight digest). the large volume is to buffer against evaporation. And don't use to much enzyme either, the glycerol that the enzyme comes in place havoc with its activity.


You can also do the digestion in your PCR cycler, which will hold the reaction at 4C after it is done. The cycler, with a hot lid, also solves the evaporation problem.