Open debate: agarose inhibition :-/

moez torki torki at NOCS.TSUKUBA-NOC.AFFRC.GO.JP
Wed Jun 9 21:30:10 EST 1999


Hi Molbiolers,

I often heard that gel purified fragments can be recalcitrant to subsequent
enzymatic treatments (eg. phosphorylation, ligation, etc..). Although I
never had problem before, now I am experiencing serious problems in my new
lab :-( That is I cannot perform even a rudimentary subcloning of my
TAE-agarose/Geneclean purified fragments. I could get rid of that problem
by phenol/chloroform/EtOHppt. I could see a whitish pellet and a white
interface after phenol extraction that I may guess being agarose ? or
silica ?
Has anyone bear such troubles ? and is there really significant differences
in agarose quality (depending on the supplier/price). Indeed I can carry on
with the phenol extraction but it doesn't really make sense since I am
using kits to get results faster...
Thanks for any comment.

Moez Torki

_________________________________________
Moez Torki, Ph.D.
Plant Biotechnology Institute,
Ibaraki Agricultural Center, 3165-1 Ago,
Iwama, Nishi-Ibaraki 319-0292, Japan

Tel:   +81-299-45-8330
Fax:   +81-299-45-8351
Email: torki at nocs.tsukuba-noc.affrc.go.jp
_________________________________________
The World : once thought to be flat, then proved to be round. Now it's
quite definitely Web shaped....





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