| 摘要 |
<p>The present invention provides a method for simultaneously determining the levels of selected target polynucleotide sequences in a sample. In the method, selected target sequences are amplified using sequence-selective primer pairs to form double-stranded copies. The copies are cleaved with one or more endonucleases, and first and second aliquots of the cleaved fragment mixture are ligated to first and second adaptors, respectively, to form mixtures of first and second adaptor-fragment conjugates. The aliquots are combined and ligated to form conjugate dimers containing a conjugate from each of the first and second conjugate mixtures. After optional amplification, the conjugates are treated to release the adaptor segments, yielding target-identifier dimers. The dimers are polymerized to form target-identifier dimer multimers, and the relative abundances of the target-identifiers in one or more dimer multimers to provide an estimate of the levels of the selected target sequences in the sample. The present invention is particularly useful for determining expression levels of mRNA gene sequences in a sample, and for evaluating changes in mRNA expression levels among different samples or in response to changes in sample conditions. Also disclosed are certain polynucleotides and polypeptides that are useful for a variety of applications, particularly relating to asthma.</p> |
