摘要 |
The present invention provides a method for quantitative and/or comparative assessment of the relative amounts of mRNA transcripts present in a cell or tissue sample. In the method reverse transcription of the mRNA contained in the sample is first carried out using sequence-modifying primers for one or several genes in the same reaction to obtain a pool of sequence-modified cDNA molecules. After completion of the reverse transcription redundant sequence-modifying primers are removed or inactivated. This step is followed by a step of co-amplifying the sequence-modified cDNA templates with a reference DNA template in individual gene-specific amplification reactions. By quantitatively measuring the amounts and determining the relative levels of the amplification products derived from sequence-modified cDNA and reference DNA templates, a gene-specific cDNA over DNA ratio is obtained in each of the individual amplification reactions. Finally, by combining the ratios obtained, a sample-specific profile can be generated, which reflects the relative amounts of mRNA transcripts originally present in the sample.
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