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Brilliant II SYBR® Green QPCR & QRT-PCR ROX Master Mixes - Details & Specifications

The Brilliant II SYBR Green QPCR Master Mix with ROX includes the components necessary to carry out QPCR amplifications with SYBR Green detection. The improved Brilliant II formulation yields higher levels of final fluorescence and earlier Ct values for many genomic DNA and cDNA targets and the master mix format is ideal for high-throughput applications. The Brilliant II master mix has been optimized for a faster two-step cycling protocol that is 25% shorter than the protocol used with the original Brilliant SYBR Green QPCR master mix. The Brilliant II master mix is also ideal for quantification of cDNA in two-step QRT-PCR reactions when combined with the AffinityScript QPCR cDNA Synthesis Kit.

The Brilliant II SYBR Green QPCR and QRT-PCR master mixes with ROX is available in two formulations: a high ROX master mix and a low ROX master mix. The final concentration of ROX reference dye in QPCR reactions prepared with the high ROX master mix is 500 nM. Reactions prepared with the low ROX master mix contain 30 nM of ROX. The two formulations are recommended for different real-time PCR platforms.

The Brilliant II SYBR Green QRT-PCR Master Mix with ROX, 1-Step kit includes the components necessary to carry out cDNA synthesis and PCR amplification in one tube and one buffer, making it ideal for most high-throughput QRT-PCR applications where it is not necessary to archive cDNA. The kit can be used to perform absolute or relative quantitation of gene expression, and has successfully been used to amplify a variety of high- and low-abundance RNA targets from experimental samples including total RNA and synthetic RNA. The kit is ideal for amplicons of up to 300 bp.

ROX dye is included in the master mix to compensate for non-PCR related variations in fluorescence. Fluorescence from ROX dye does not change during the course of the PCR reaction but provides a stable baseline to which samples are normalized. In this way, ROX detection compensates for changes in fluorescence between wells caused by slight volume differences in reaction tubes. The excitation and emission wavelengths of ROX dye are 584 nm and 612 nm, respectively.

In addition to ROX, the QPCR master mix includes SureStart Taq DNA polymerase, nucleotides (GATC), and an optimized QPCR buffer. SureStart Taq DNA polymerase, a modified version of Taq2000 DNA polymerase with hot start capability, improves PCR amplification reactions by decreasing background from nonspecific amplification and increasing amplification of desired products. Using SureStart Taq, hot start is easily incorporated into PCR protocols already optimized with Taq DNA polymerase, with little modification of cycling parameters or conditions.

For Research Use Only. Not for use in diagnostic procedures