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Old 07-06-2011, 09:04 AM   #2
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Location: Purdue University, West Lafayette, Indiana

Join Date: Aug 2008
Posts: 2,317

If I remember correctly, T4 PNK will add a phosphate to a hydroxyl 5' end and/or remove a phosphate from the 3' end of a DNA or RNA strand. This is the desired phosphate configuration for most ligases.

What is the end produced by the enzyme being used to fragment the library? Typically a restriction enzyme will give 5'-phosphate/3'-hydroxyl that is needed. But the overhangs may or may not be compatible.

You can get the number of units necessary per mole of your fragment ends from the web site of the manufacturer you order the enzyme from.

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