AnaSpec offers a high-throughput platform for producing small-scale real-time qPCR double-dye probes for genotyping or gene expression profiling, requiring small amounts of several diffent probes.
Quality controlled by analytical HPLC and MALDI-TOF Mass Spectrometry, the probes are provided lyophilised in individual tubes.
Double-dye oligonucleotides have a fluorescent reporter dye and a quencher at their 5' and 3' ends.
These oligos are designed to anneal between the upstream and downstream PCR primers.
The higher Tm of the double-dye oligonucleotide probe compared to the Tm of the upstream primer ensures that the primer is already being extended while the double-dye oligonucleotide probe is not yet annealed.
The optimal recommended length of double-dye oligonucleotide probes is 25 bases, maximum length 30 bases.
For extra-long double-dye oligonucleotides probes, quenchers like Black Hole Quencher, Deep Dark Quencher I and TAMRA can be coupled internally.
