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DNA preparation for microinjection - (Apr/03/2005 )  Hello
everyone,  I have some questions regarding the preparation of
DNA for Tg mouse generation. I haven't done this before and have
some problems regarding the quantity of the DNA. I use 50ug of
plasmid DNA for restriction digestion and after gel purification
of the fragment, there is only ~300ng left. The fragment is not
small (7kb of a 10kb plasmid) and I don't really understand
where is it lost. Is that normal? After cutting, I use the
Qiagen PCR Purification Kit and then gel purify (Qiagen Gel Extr
kit).  Also is that the correct way to submit a DNA fragment for
microinjection. Just cut out and purify. I mean shouldn't I
blunt-end the overhangs or something?  Thanks really for any
suggestions.  Best Regards to All,    ok, firstly, I just wanna
make sure that you used 50 micrograms of DNA. That is a LOT of
DNA.  Secondly, What is the capacity of the columns in your
Qiagen kits?   Thank you