Good poly-A+ isolation kit, anyone???

Karthik Aghoram kaghoram at unity.ncsu.edu
Tue Aug 8 08:19:52 EST 2000


I have used only one ... Dynal's Dynabeads; and they worked like a 
charm.  If you have the $$$ or ###, you should try it.

Good luck.
Karthik (no affiliation to Dynal Inc)


Mr B.J.H. Jansen (bjansen at hgmp.mrc.ac.uk) 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
: 
: Bas
: 
: Bastiaan JH Jansen
: Dept of Dermatology 802
: University Medical Centre St Radboud
: PO Box 9101
: 6500 HB Nijmegen
: The Netherlands
: Tel: +31 (0)24 3617239
: Fax: +31 (0)24 3541184
: Email: basjhj at mailbox.kun.nl
: 

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Karthik Aghoram				Postdoctoral Associate
					Dept of Crop Science
e-mail: kaghoram at unity.ncsu.edu		North Carolina St University
Phone (W): 919-515-2705			Raleigh, NC 27695-7620

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