cDNA kit recommendations?

Michael L. Sullivan mlsulliv at facstaff.wisc.edu
Mon Mar 26 12:49:26 EST 2001


Been there, done that.  In the end it's sort of fun knowing you have the
only cDNA library in the world for something!

I made a cDNA library about a year ago using a kit from Stratagene (ZAP
cDNA synthesis kit) [I have no affiliation with Stratagene].  I have a lot
of molecular experience but I'd not actually ever made a library.  I think
the Strategen kit is very straightforward and fairly fool proof.  One
reason I chose it was I was fairly familiar with Strategene's Lambda Zap
vector, which can be rescued to a pBluescript phagemid.  Overall, a fairly
conveinient system in my opinion.

The only thing I did different from the kit, was  the size fractionation
step, which is fairly cumbersome, and results in your cDNA being diluted
among several fractions.  Somebody in this forum gave me the tip to use a
spin column product from Pharmacia to do the size fractionation (SizeSep
400) [I've no affilitation with Pharmacia either!].  I did this, and it was
much easier than the column proceedure Stratagen included would have been
and worked very well.  I found a few tricks to using the Pharmacia spin
column that I can share with you if you go that route.

These two products worked very well for me and if I were doing it again,
I'd make the same choice.  If you'd like any more input, feel free to let
me know.  I'd be happy to discuss it more with you.

Mike

>I need to make a cDNA library from a novel plant species
>(i.e. NO pre-made libraries exist, alas!).  I have not in
>fact ever actually made a library before and I was wondering
>if people had recommendations for which kits to use/not use?
>I am particularly interested in any brands which I should
>avoid, as I'd rather not deal with poor product in addition
>to my own inexperience.  Any advice is appreciated!
>
>--
>Allison Mackay
>Plant Cell Technology Laboratory
>Department of Plant Agriculture
>University of Guelph
>
>(519) 763-7165
>*** send attachments to deep_fried_gmos at yahoo.ca ***


Michael L. Sullivan, Ph.D

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

(608) 264-5144 Phone
(608) 264-5147 Fax

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