before electro-transformation, remove T4 Ligase or not? - (Oct/26/2008 )

I have searched the Forum (maybe not long enough) and i did not get an answer. I have not much experience in molekular biologie, so my question is: Do I remove the T4 Ligase before i do the electro-transformation into e.coli? If the answer is yes. How do i remove/inactivate T4?
Thanks for your help.

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You have to desalt your ligation mix.. and this means you have to eliminate the salt in the ligation buffer (not ligase itself). Otherwise you'll get a spark in the electroporation cell, which will be shown in Err or something like that in your electroporator.

To desalt your ligation mix, there are many methods including ethanol precipitation, and column purification. Google it.

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No. You don't have to remove the T4 ligase. However as already stated if you use electroporation, you have to desalt your ligation mix. I prefer ethanol percipitation followed by large 70% ethanol wash (~200ul) as it is cheaper. However you must be careful not to lose the DNA pellet.

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Yeah, I also didn't desalt the ligation mixture at all but it still worked well. Only 1 time when I transform, the "Arc" error appeared, maybe due to the high volume of ligation mixture I used at this time.

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