Nucleic Acids Research, Vol 25, Issue 22 4694-4696, Copyright © 1997 by Oxford University Press
K Kato
A simple and reliable PCR-based method to quantitate gene expression is
described. Following the digestion of double-stranded cDNA by a restriction
enzyme, an adaptor is ligated to a cDNA from a first RNA sample, and
another adaptor to a second RNA sample. The two adaptors share a common
sequence at the outer region, but differ in size. Equal amounts of the
ligated samples are mixed, and amplified by an adaptor- primer and a primer
specific to the gene of interest. Products derived from the two sources
differ in size, and can be separated by denaturing polyacrylamide gel
electrophoresis. The ratio of the two products reveals the relative level
of gene expression. Since the technique avoids the need to construct
internal standards, it is especially useful for the analysis of many
different gene transcripts.
ARTICLES
Adaptor-tagged competitive PCR: a novel method for measuring relative gene expression
Louis Pasteur Center for Medical Research and Cell Switching Project, ERATO, JST, 103-5 Tanakamonzencho, Sakyo-ku, Kyoto 606, Japan. k.kato@bs.aist-nara.ac.jp
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