PCR Ligation Issues!!!

ChenHA hzhen at freeuk.com
Wed Sep 28 11:53:00 EST 2005


On Tue, 27 Sep 2005 23:54:57 +0100, Peter Ellis <pjie2 at cam.ac.uk>
wrote:

>Why are you purifying it on a gel?  If you do a column prep, you won't 
>need to nuke it with UV.
>

There are good reasons why a gel purification is often better than
using column.  I would nearly always include a gel purification step
in cloning, for various reasons, for example, if the vector used for
the PCR has the same restriction site (so a gel purification step
after PCR), or  if the digested fragments you want to remvoe is too
big to be effectively removed by column.   No problem with UV either,
as long as you use the right equipment, although  the original poster
use crystal violet and don't need to use UV.



>Peter



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