Abstract: The present invention is directed to methods, reagents, kits, and compositions for identifying and quantifying target polynucleotide sequences. A linker probe comprising a 3? target specific portion, a loop, and a stem is hybridized to a target polynucleotide and extended to form a reaction product that includes a reverse primer portion and the stem nucleotides. A detector probe, a specific forward primer, and a reverse primer can be employed in an amplification reaction wherein the detector probe can detect the amplified target polynucleotide based on the stem nucleotides introduced by the linker probe. In some embodiments a plurality of short miRNAs are queried with a plurality of linker probes, wherein the linker probes all comprise a universal reverse primer portion a different 3? target specific portion and different stems. The plurality of queried miRNAs can then be decoded in a plurality of amplification reactions.

Abstract: Compositions, methods, and kits for identifying and for quantitating polynucleotide targets are disclosed. These compositions, methods, and kits are particularly useful when the polynucleotide target is a small RNA molecule, including without limitation microRNA (miRNA), small interfering RNA (siRNA), and certain other classes of non-coding RNA molecules. The forward and reverse primers of the disclosed first primer sets comprise unusually short target-binding portions that are 6-10 nucleotides long. Certain of the disclosed methods employ one or more multiplex reaction steps to identify, quantitate, or identify and quantitate, a multiplicity of target polynucleotides.

Abstract: The present invention relates to the detection of target sequences. The present description discloses compositions and methods involving analog nucleic acids, such as PNA and L-DNA, for the detection of nucleic acids. Additionally, hybrid detectable markers are provided.

Abstract: A simplified method for isolating primer extension products and generating them in a form appropriate for electrophoresis is disclosed. The method is compatible with automated DNA sequencing procedures.