SsoFast™ EvaGreen® Supermix: Life Science Research: Bio-Rad

SsoFast™ EvaGreen^® Supermix

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Overview

Sso7d Fusion Protein Technology

Bio-Rad’s patented* Sso7d fusion protein technology was first employed in the iProof™ high-fidelity DNA polymerase product line. The unique Sso7d fusion polymerase in the SsoFast™ supermixes enables fast cycling without affecting PCR sensitivity, efficiency, and reproducibility. The dsDNA-binding protein, Sso7d, stabilizes the polymerase-template complex, increases processivity, and provides greater speed and reduced reaction times compared to conventional DNA polymerases. Furthermore, this polymerase is significantly more resistant to PCR inhibitors, making the SsoFast supermixes ideal choices for applications such as direct qPCR, without the need for sample preparation.

Fusing the double-stranded DNA binding protein Sso7d to DNA polymerases provides a powerful sliding grip on the replicated DNA.

* U.S. patents 6,627,424; 7,541,170; and 7,560,260.

Performance

The unique fusion polymerase in SsoFast EvaGreen supermix delivers extreme speed and generates exceptional qPCR results in less than 30 min. 10-fold serial dilutions of 10 ng to 100 ag of cDNA from human spleen were used in each 20 µl reaction to detect 18S RNA. 18S efficiency = 101.8%, r = 0.997. Total qPCR run time = 29 min. RFU, relative fluorescence units.

Sensitivity

SsoFast EvaGreen supermix provides extreme sensitivity in detection of a single copy of target gene. The ZAP70 gene was amplified and detected from 5-fold serial dilutions of 10 ng to 80 pg () and 3.2 pg () of human genomic DNA. ZAP70 efficiency = 102.7%, r = 0.991. Insert shows the standard curve for the various dilutions. RFU, relative fluorescence units.

Reproducibility

Exceptional reproducibility can be achieved with SsoFast EvaGreen supermix. Efficient discrimination and reliable quantification can be obtained from 1.33-fold serial dilutions of input template. The CBP gene was amplified from varying amounts of human genomic DNA (5 ng to 500 pg). From left to right: () 5 ng, 2.83 ng, 1.60 ng, 903 pg, and 511 pg; () 3.76 ng, 2.13 ng, 1.20 ng, and 679 pg. CBP efficiency = 96.5%, r = 0.996. Insert is a magnified view showing robust discrimination and reproducible amplification. RFU, relative fluorescence units.

Bio-Rad introduces the SsoFast™ EvaGreen^® supermix, a member of our next generation of real-time PCR supermixes using our patented Sso7d fusion protein technology, delivering a reagent that provides effective performance in a wide range of qPCR applications. The dsDNA-binding protein, Sso7d, stabilizes the polymerase-template complex, increases processivity, and provides greater speed and reduced reaction times compared to conventional DNA polymerases, without affecting PCR sensitivity, efficiency, or reproducibility.

Key Features and Benefits

Fast qPCR results and high performance — the Sso7d fusion polymerase and optimized buffer deliver fast reaction times via instant polymerase activation and rapid polymerization kinetics to generate exceptional qPCR results in less than 30 min
Minimal inhibition of PCR — the polymerase's increased resistance to PCR inhibitors ensures maximum efficiency, sensitivity, and reproducibility, and provides increased fluorescence compared to SYBR^® Green
Single copy detection — data illustrate high sensitivity with amplification and detection from a single copy of target gene
Robust discrimination and reproducibility — efficient discrimination and reliable quantification can be obtained from 1.33-fold serial dilutions of input template

Applications and Uses

Dye-based qPCR detection
PCR and real-time PCR analysis
Nucleic acid amplification and expression profiling
Gene expression analysis
Genotyping studies
Genomics-related applications

Product expiration (after ship date)	12 months
Storage at +4°C (after thawing)	Up to 6 months
Customer guarantee (minimum usage time)	12 months