Conjugates between a minor groove binding molecule, such as the trimer of
1,2-dihydro-(3H)-pyrrolo[3,2-e]indole-7-carboxylate (CDPI.sub.3), and an
oligonucleotide form unusually stable hybrids with complementary target
sequences, in which the tethered CDPI.sub.3 group resides in the minor
groove of the duplex. These conjugates can be used as probes and primers.
Due to their unusually high binding affinity, conjugates as short as
8-mers can be used as amplification primers with high specificity and
efficiency. MGB conjugation also increases the discriminatory power of
short oligonucleotides, providing enhanced detection of nucleotide
sequence mismatches by short oligonucleotides. The MGB-conjugated probes
and primers described herein facilitate various analytic and diagnostic
procedures, such as amplification reactions, PCR, detection of
single-nucleotide polymorphisms, gene hunting, differential display,
fluorescence energy transfer, hydrolyzable probe assays and others; by
allowing the use of shorter oligonucleotides, which have higher
specificity and better discriminatory power.