so many claims for so many kits -- how can we tell what works?

Jon Nakamoto jnakamot at ucla.edu
Thu Dec 7 07:09:42 EST 1995


   Every day several ads for appealing-sounding kits cross my desk
(generally selling for several hundred dollars a pop). I keep an eye out
for postings describing these kits, but see surprisingly little discussion
about them. Surely there are people out there who've tried these kits
firsthand and can share their acquired wisdom?
   I'm not looking for full-fledged reviews, nor would I want to see bad
mouthing of any specific product. Just some reality checks and tips about
specific kits. 
   For example, I've had the chance to try the Gibco BRL Gene Trapper Kit,
which basically uses a biotinylated oligo and streptavidin-paramagnetic
beads to enrich for the desired clones. It does work, but I would comment
that you pull down a lot of clones which have one or two mismatches from
the oligo sequence (and that means you could get a lot of clones other
than the one you're after). Also, the TdT/biotin-14-dCTP incorporation
step is really fussy and demands a PAGE-purified oligo, even though the
protocol says you can use HPLC-purified oligos -- if someone had told me
this from the beginning, it would have saved me a lot of grief (and
reagents). Also, it's important to note that an extremely high efficiency
of bacterial transformation is essential for this kit -- you have to use
electroporation. 
   Another example: an experienced colleague tried the Clontech Promoter
Finder kit to get very G+C-rich 5' sequence but had little luck. It would
be nice to hear from someone who had been successful in getting GC-rich
sequences with this kit, or others who could confirm that it was likely
the GC-richness and not technical incompetence on my colleagues part! 
   I'm not trying to push kits (hey, I set up my own differential display
of mRNA rather than paying the astronomical GenHunter price), but
sometimes they do work nicely and save much time. I just want to know
which ones have worked well for your average lab person, and just what
that average person did to get it working. 
   Why isn't there a site where you can look up specific products and get
users' tips for how to make a particular product work best? 

Sorry for the late night rambling,
Jon

Jon Nakamoto, MD
UCLA Medical Center
jnakamot at ucla.edu

-- 
Jon Nakamoto
jnakamot at ucla.edu



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