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SYBR® Green This dye preferentially binds to double-stranded DNA and results in a strong fluorescence emission signal. The intensity of this signal is proportional to the amount of double-stranded DNA present; therefore this dye can be utilized to monitor the accumulation of amplified products resulting from qPCR amplification (Figure 1A). SYBR® Green is widely used both because of its high detection sensitivity and its relatively low cost. This dye does require high-quality primer designs to avoid primer dimers that could cause non-specific increases in the signal. Advances in the understanding of the PCR process and the developmentof sophisticated primer design algorithms have greatly helped in addressing this issue. Under carefully controlled conditions, SYBR® Green PCR works extremely well. SABiosciences Corporation leads the field of SYBR® Green PCR, featuring advanced SYBR® Green technologies including PCR Arrays, the unique hotstart Taq polymerase, and > 60,000 genome-wide certified primer assays.

DNA Hydrolysis Probes Another major detection method for PCR-based amplification is based on the 5'to 3' exonuclease activity of Taq polymerase. This exonuclease activity is used to cleave a fluorogenic hybridization probe - TaqMan® Probe - which consists of an oligonucleotide recognizing an internal region of the target sequence. Theprobe is labeled with a 5' fluorescent reporter dye and a 3′quencher dye in closeproximity. A signal is generated when the Taq polymerase hydrolyzes the 5' endof the probe while polymerizing the target DNA, and releases the reporter dyefrom the quencher (Figure 1B). The primary advantage of the DNA hydrolysis PCR assay is that it will not detect non-specific products such as primer dimers. Adrawback is that it requires a specific probe for every gene target, thereby increasing the cost. In 2008, SABiosciences introduced the first cost-effective DNA Hydrolysis Array solution featuring this technology and optimized Probe Master Mixes for all RT-PCR instruments.

Unique Elements of SABiosciences qPCR Master Mixes
Master Mix quality plays a critical role in the performance of real-time PCR. The key to obtaining accurate results is a tightly-controlled hot start enzyme that eliminates the amplification of primer dimers and other secondary products. Antibody-based technologies have worked well in the past for most PCR applications. However, these enzymes too often "leak" and synthesize too many non-specific amplicons to meet the more rigorous and demanding standards set by SABiosciences for real-time detection of message transcripts. After years of experience in designing and experimentally testing thousands of PCR primers for entire genomes, SABiosciences has developed a qPCR master mix formulation that provides the most accurate real-time PCR results.

Unique HotStart Polymerase and Buffer Components
The RT² qPCR Master Mixes from SABiosciences offer superior specificity and amplification efficiency thanks to our high-quality chemically-modified HotStart Taq DNA polymerase. Hot start enzymes found in other sources of master mix, such as the antibody-based technologies, still retain residual activity in their "inactive" state at low temperature. Under these conditions, non-specific priming events are extended to form templates which are then amplified into non-specific products that artificially contribute to signal intensity, resulting in unnecessary false positive results. Our tightly controlled HotStart enzyme lacks any activity before being heated and therefore minimizes the amplification of non-specific PCR products.

The RT² qPCR Master Mix formulations also include other proprietary chemical components that significantly minimize primer dimer formation ensuring high amplification efficiencies for even the most difficult to amplify genes.

All-in-One Master Mixes: Instrument Reference Dyes Included
Our RT² qPCR master mixes are tailor-made and optimized for the specific real time system in your lab. These cost-effective all-in-one PCR master mixes not only contain optimized buffers, nucleotides, and our high-quality HotStart DNA polymerase, but they also include the appropriate reference dye specifically used by individual real time systems to normalize their optics. Just add your primer pairs and templates, and you're ready for PCR.

Complete your gene expression experiments with RT² qPCR Primer Assays, RT² Profiler PCR Arrays, or other commercially available gene expression assays with the appropriate RT² qPCR Master Mix tailor-made to the instrument in your laboratory.

SYBR® is a registered trademark of Molecular Probes. ABI® is a registered trademark of Applied Biosystems. iCycler® is a registered trademark of BioRad Laboratories, Inc. SmartCycler® is a registered trademark of Cepheid Purchase of this product includes an immunity from suit under patents specified in the product insert to use only the amount purchased for the purchasers own internal research. No other patent rights are conveyed expressly, by implication, or by estoppel. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.