The present invention relates to a method for generating transcripts from:
at least one RNA sequence to be amplified comprising a "primer" region
and a region of interest, and an amplification primer comprising a
promoter region, and a region capable of hybridizing to said "primer"
region of the RNA sequence to be amplified, said method being carried out
at constant temperature, and comprising the following steps: a) said
primer is hybridized with the RNA to be amplified, b) the primer is
extended by means of a reverse transcriptase enzymatic activity in order
to generate a complementary deoxyribonucleic acid (cDNA) sequence of the
RNA to be amplified, c) the RNA to be amplified, hybridized to said cDNA,
is cleaved by means of an enzyme that has a ribonuclease H activity, so
as to obtain fragments of RNA to be amplified, hybridized to said cDNA,
d) the ends of said fragments of RNA to be amplified are extended by
means of a reverse transcriptase and strand displacement enzyme, so as to
obtain RNA-DNA/DNA hybrids, e) RNA transcripts are obtained from the
RNA-DNA/DNA hybrids formed in step d), by means of an enzyme that has an
RNA polymerase activity.