Clontech's SMART - reference please?

Alexander S Kraev alexander.kraev at utoronto.ca
Fri Jan 28 16:40:06 EST 2000


Dear netters,
I have recently come across a particularly difficult case of locating  a
5'-end of an RNA,
which prompted to search for alternatives for doing the RACE experiments. I
have been
using a modification of the original procedure, that is tailing with deoxy-A
and TdT, then
making second strand with an anchor primer, ending in oligoT, and finally
amplifying the
product using PCR. In most of the resultant products  sequenced with dye
terminator
chemistry directly the dT tail is not attached right to the presumable cDNA
end but
via three non-complementary G (on the coding strand) residues.  Now, I  have
come across a Clontech
kit which claims that (citation from the page 50 of the year 2000 catalog)
"when reverse
transcriptase reaches the end of the mRNA template, it adds a few (dC)
residues...". Could
anybody, i.e. from Clontech or other please provide a reference to the paper
describing the
primary data behind the (apparently) patented SMART technology? Are we led
to believe that
the three C residues I am seeing with Expand RT added to the cDNA end are
the basis of SMART?

This question is so vital to me because I believe to be isolating a product
of a cDNA synthesis block rather than a true capped 5'-end.

Alex




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