so many claims for so many kits -- how can we tell what works?
Kay A. Pleyte
kpleyte at post.its.mcw.edu
Thu Dec 7 11:25:00 EST 1995
I've found lots of comments on various kits by searching the Usenet
archives. I agree the comments of others are the most useful evaluation of
a new product's performance in "real life". It takes a while for people to
try a new product. I always wait for other people to try before I take the
chance! So, keep the new product evaluations coming--they're probably read
by a lot of researchers.
KP
kpleyte at post.its.mcw.edu
Jon Nakamoto (jnakamot at ucla.edu) wrote:
: 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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