Good poly-A+ isolation kit, anyone???
patrick.merel at chu-bordeaux.fr
Wed Aug 9 10:48:22 EST 2000
Paraic Kenny wrote:
> Hi Bas
> I've used both Fasttrack from Invitrogen and Quickprep from Pharmacia
> with variable success (FastCRAP, and pretty OK respectively) but now I
> use Oligotex from Qiagen and it works really well in my hands. Very fast
> protocol, and with great yield and A260/280 and no ribosomal bands. A
> bit more expensive, but worth it for the end product.
> Good luck
> "Mr B.J.H. Jansen" wrote:
> > Dear BioNetters,
> > Currently I'm in the process of constructing some good old c-DNA libraries.
> > The input m-RNA comes from cultured keratinocytes, and the problem is that
> > these do not grow to high density (in Keratinocyte Growth Medium up to 5*10^5
> > cells per standard dish of 20 cm^2), which means that I've to use an awful
> > lot of lysis buffer because of the large surface area on which these cells
> > grow. I have to isolate total RNA first, and yields are fairly consistent.
> > So far I have used kits from Pharmacia and Roche to further purify poly-A+
> > RNA, with widely varying results. What bugs me is the 'widely varying'part,
> > and I was wondering whether one of you has ever compared various poly-A+
> > isolation kits. I have so far isolated poly-A+ from 50 to 100 micrograms of
> > total RNA, admittedly not very much, but in one case it has already resulted
> > in the birth of a c-DNA library. Oh yeah, since we try to pick up genes that
> > have been identified in SAGE libraries, commercially available c-DNA libraries
> > are not an option. Thank you for your help!
> > All the best
give a try to RNAture.com
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