mRNA purification

eric anderson e-anderson at ski.mskcc.org
Tue Dec 3 15:16:28 EST 1996


In article <32a48152.0 at robby.oit.umass.edu>, Mike Holbrook
<holbrook at mcb.umass.edu> wrote:

> I sure this subject has been addressed in this newsgroup, but I missed 
> the thread.  Of the kits available for total mRNA isolation, which has 
                                         ^^^^^^^^^^^^^^^^^^^^^
<snip>
> All assistance is greatly appreciated.
> 

mike,

do you want total RNA isolation or do you want Poly-A++ mRNA isolation?

for the first, I would suggest one of the TRI reagents, lots of companies
make them, we happen to use Sigma's (#T9424, 200ml, $165).  It takes about
1ml of reagent to treat .5-1x10^7 suspension cells or 1 10cm^2 dish.  this
works every time, is very simple and takes about 2.5 hours start to finish
for 4 samples isolated at the same time (YMMV).  hands on time is about
45-60 min.

for poly-A++ mRNA (from total RNA) i would try using the Promega
PolyATract kit.  for isolation of poly-A++ mRNA directly from cells, try
the Qiagen Oligotex Direct kits.  i've had good luck with both of these
kits (although not with Qiagens total RNA isolation kits) and recently
chose to stick with the Promega kit because it's relatively inexpensive
and gives acceptable yields.
the Invitrogen FastTrack kits work well also but they're very expensive
and kind of a pain to use compared to the Promega and Qiagen kits.

good luck,

eric

-- 
Eric C. Anderson
Memorial Sloan-Kettering Cancer Center
Sloan-Kettering Institute
1275 York Ave. Box 470
New York, NY  10028
(212) 639-2977
e-anderson at ski.mskcc.org



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