Patent ID: 7838228
Filing Date: 2010-11-23
Classification: C12Q

Abstract:
1. A method for quantitative and/or comparative assessment of the relative amounts of mRNA transcripts present in a cell or tissue sample, the method comprising the steps of: a) carrying out reverse transcription of the mRNA contained in the sample using sequence-modifying primers for several genes in the same reaction to obtain a pool of sequence-modified cDNA molecules, wherein the sequence-modifying primers are oligonucleotides comprising three functional segments: b) removing or inactivating redundant sequence-modifying primers after completion of the reverse transcription, c) carrying out individual amplification reactions for each of the analyzed genes, so that in each of the reactions sequence-modified cDNA templates are co-amplified with a reference DNA template, using gene-specific primers to generate measurable amounts of amplification products, wherein the gene-specific primers have binding sites which are located on the same exon, d) quantitatively measuring the amounts and determining the relative levels of the amplification products derived from sequence-modified cDNA and reference DNA templates to obtain a gene-specific cDNA over DNA ratio in each of the individual amplification reactions, and e) combining the gene-specific cDNA over DNA ratios determined in the individual amplification reactions to generate a sample-specific profile of said ratios that reflects the relative amounts of mRNA transcripts originally present in the sample.