cDNA help need

Hallick Lab hallick_lab at TIKAL.BIOSCI.ARIZONA.EDU
Fri Dec 9 12:01:42 EST 1994


        Reply to:   RE>>cDNA help need

ATA makes the RNA useless for anything but northerns, definitely get rid of
it.  I tried using it many years ago when making tassle-specific cDNA
libraries from corn.  Nothing worked until I left out the ATA,  then it
cloned like gangbusters
Mitch Favreau  

--------------------------------------
Date: 12/9/94 7:15 AM
To: Hallick Lab
From: Hanson Lab
In article <D0H8MB.IJq at osuunx.ucc.okstate.edu>, Wenjun Huang
<biocwjh at osuvm1.bitnet> wrote:

> 	I need a help! (for my first strand cDNA synthesis from cotton cell
> suspension culture)
[a big deletion of the description of the prep]
Wenjun,
I know how frustrating it is to have cDNA synthesis not work.  And I work
with plant material.  I have a few suggestions.
-I am not sure, but I thought ATA inactivated RNA as well as the RNases
that you add it for.  I would try leaving it out of your prep.
-Better though, I suggest trying the RNA prep described for plant material
in Current Protocols (the Red Book).  It has worked well for me.  There are
other RNA preps that might work as well (all of the "new" Guanidine-phenol
types) but I have no experience with them.
 Good luck, in any case.
Claudia Sutton  aka cas9 at cornell.edu


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