( b (Figure 2 ). This large dyn...)Brilliant Core Reagent Kits for QPCR and QRT-PCR

b> (Figure 2). This large dynamic range eliminated the need to analyze serial dilutions of experimental samples for gene quantification. Moreover, this target titration demonstrated that the Brilliant Plus QPCR kit is sensitive enough to detect as few as 10 copies of plasmid DNA.

To test the compatibility of the Brilliant Plus QPCR core reagent kit with different fluorescent chemistries, we used a Scorpions probe to detect and amplify the 134-bp rpoB gene target of Mycobacterium tuberculosis, an emergent pathogen. We amplified and detected 100, 1000, and 10,000 copies of the M. tuberculosis target gene; the respective Ct values were 38.2, 34.9, and 31.6 (data not shown).

Multiplex Capabilities

Fig.3

In further experiments, we showed that the Brilliant Plus QPCR reagent kits can efficiently detect more than one dye per reaction (Multiplex QPCR or MxQPCR). We used Stratagenes molecular beacon SDF-1 (Stromal Derived Factor 1) Allelic Discrimination kit, which detects a common G-to-A substitution that may delay or accelerate the onset of AIDS in humans (Figure 3). We detected this substitution by amplifying human genomic DNA in the presence of two molecular beacons in the same tube, each molecular beacon with a different dye (FAM and TET). In this scheme, the FAM probe was complementary to the mutant allele (A), and the TET probe was complementary to the wild-type allele (G). We included the passive reference dye, although not required, as the third fluorophore. The genotype-specific control DNA samples supplied in the molecular beacon SDF-1 kit were used for this allelic discrimination.

This multiplex assay provides a way of determining whether a
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