详细信息:The Amplifluor Mouse Nanog JOE real time PCR primer set was designed for the quantitative evaluation of Nanog gene expression in mouse ES and iPS cells. It can be used in either single gene detection assay or together with other fluorophore conjugated primer sets in multiplex detection assays. Assay conditions may need to be further optimized for multiplex detection.
This kit utilizes the Amplifluor system for the detection of RNA expression levels. The Amplifluor RT-PCR reaction is performed using a reverse transcription (RT) primer for 1st strand cDNA synthesis, followed by Polymerase Chain Reaction (PCR) amplification utilizing the forward Amplifluor hairpin primer and reverse primer to produce gene-specific fluorescent amplicons. The Amplifluor hairpin primer is labeled with Donor (fluorophore) and Quencher (DABSYL), which produce the high signal to noise ratio observed in the Amplifluor assay. Upon incorporation into new amplicon, the Amplifluor primer is fluorescent, while the fluorophore signal of the unincorporated Amplifluor primer is quenched. When performed on a thermal cycler that is capable of real-time fluorescence detection, the amplification products of the PCR reaction can be monitored directly in a quantitative fashion, allowing evaluation of template detection in a single, closed-tube system with no associated assay procedure. Where required, template standard curves may be employed to quantitatively determine sample load.
