request advice on mRNA preparation

Klaus Salger salger at wap18.zi.biologie.uni-muenchen.de
Sat Oct 29 15:36:39 EST 1994


Robert Hewitt (rhewitt at HELIX.NIH.GOV) wrote:
: Dear Internetter,

: I've been using a combination of the Stratagene micro RNA isolation 
: kit and the 5'-3' spin column poly A selection kit, to make polyA RNA 
: from human colon cancer cell lines.  My yields of mRNA are only 1-2% of 
: the total starting RNA.  I'm told by the 5'-3' technical advisers, that 
: these yields are on the low end of what you would expect for cell line 
: RNA, and that you get higher yields with tissue RNA.  They also advise 
: that you get higher yields if you poly A select from freshly prepared total 
: RNA.  Can anyone suggest why this is?  Any comments or suggestions would 
: be greatly appreciated. Thank you in advance,

: Robert Hewitt

Dear Robert,
1-2% mRNA is what you expect according to Sambrook et al.

Klaus

--
Klaus Salger                phone : ++49 (0)89 5902 -502
Zoologisches Institut       FAX   :                 -450
AG MacWilliams              e-mail: salger at zi.biologie.uni-muenchen.de
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