Looking for good RNA extraction columns

Michael L. Sullivan mlsulliv at facstaff.wisc.edu
Wed May 17 08:58:37 EST 2000


I personally have had really good success with the RNeasy columns from
Qiagen.  I got a reasonable amount of RNA that really looked nice on a gel
(and eventually made a nice cDNA library).  My biggest problem with them
was that they are designed for pretty small scale isolations (like from 100
mg of tissue), so I ended up using several columns to get the amount of RNA
I needed to make poly(A) RNA.  On the plus side though, they are easy,
fast, not horribly expensive, and given your insecurity about getting stuff
RNAse free, virtually everything (well, not pipette tips) is provided
including collection tubes.

Good luck

Mike



>      We are attempting to extract total RNA from
>soybean roots, epicotyls, hypocotyls, and
>cotyledons, with the eventual intent of running
>RT-PCR and Northern analysis.  We've tried twice
>now with the Chomczynski & Sacchi procedure
>(flash-freeze in liquid Nitrogen, GTC, phenol/
>ChCl3 extractions, EtOH washes, etc etc etc).
>Each time, after we run it out on the formaldehyde
>gel, all we see are smears in the RNA lanes, with
>no real discernable bands.  We suspect severe
>degredation somewhere along the way.  Our only
>real prevention measures are to keep a seperate
>set of tips and reagents just for RNA use, and to
>RNAseOUT surfaces, pipetors, and gloves.  However,
>due to the layout of the equipment in the
>department, we currently have to move between
>several rooms in order to complete the
>extractions, and we can't just RNAseOUT the entire
>world.  We'd like to try using columns or a kit
>that would help reduce the number of steps
>involved and, subsequently, the potential for
>RNAse contamination.  I've read a lot about QIAgen
>RNeasy kits, but I've also read where people sing
>the praises of Trizol.  Does anybody have any good
>suggestions of alternative protocols or kits to
>use or, perhaps more valuable, ones to stay the
>heck away from?  Also, are Ambion's RNAse-free
>tubes and tips worth the extra cash, or can you
>just do as much with autoclaving and segregating
>your materials?  Thank you very much.
>
>Adam R. Hoagland
>Terzaghi lab
>Wilkes University
>Wilkes-Barre, PA
>USA
>
>
>Sent via Deja.com http://www.deja.com/
>Before you buy.


Michael L. Sullivan, Ph.D

U.S. Dairy Forage Research Center
1925 Linden Drive West
Madison WI, 53706

(608) 264-5144 Phone
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