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USB® Hot Start qPCR products--consistency, value, and performance
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Consistency you can count on
USB® HotStart-IT® qPCR master mixes provide the consistency found in other popular mixes such as ABI TaqMan® Universal PCR Master Mix (Fig. 1 and 2). Both mixes provide efficient amplification and target detection. In addition, USB® HotStart-IT® Taq DNA Polymerase requires only a 2 minute heat activation step compared to upwards of 10 minutes for AmpliTaq Gold®*. |
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Figure 1. Comparison of USB® HotStart-IT® Probe qPCR master mix vs. ABI TaqMan® Universal PCR master mix. TaqMan® assay under standard ABI cycling conditions to quantify expression of reference gene. All reactions were performed in duplicate. Data courtesy of Anne Jedlicka, Johns Hopkins Malaria Research Institute, Baltimore, MD. |
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Figure 2. Comparison of USB HotStart-IT® Probe qPCR Master Mix vs. ABI TaqMan® Universal PCR master mix. TaqMan® assay under standard ABI cycling conditions to quantify expression of gene of interest in both control (PBS) and experimental (RWP) samples. All reactions were performed at least in triplicate, except for IFNGKO_PBS control. Bars indicate standard deviation. Data courtesy of Anne Jedlicka, Johns Hopkins Malaria Research Institute, Baltimore, MD. |
Unmatched value
Compare the price of USB® HotStart-IT® Probe qPCR Master Mix to ABI TaqMan® Universal Master Mix. USB® HotStart-IT® provides reliable real-time PCR amplification at a lower cost per reaction. | |
Reliable performance
USB® HotStart-IT® qPCR products utilize a novel primer binding protein to inhibit primer dimer formation. The result is sensitive and consistent amplification for real-time PCR. USB® HotStart-IT® qPCR master mixes deliver sensitive detection of single copy targets and are functionally tested to ensure a linear dynamic range over seven orders of magnitude (Figure 1). Master mixes are available with SYBR® Green or without for use with fluorescent probes (e.g. TaqMan®). |
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Figure 3: GAPDH Assay using USB® HotStart-IT® SYBR® Green qPCR Master Mix (PN 75762). Triplicate reactions were performed with a cloned region of the human GAPDH gene as template using an ABI 7500 Fast instrument. The non-specific dsDNA binding dye, SYBR® Green I, was used to detect the 122 bp amplicon and ROX was used as a passive reference dye. The amplification process was linear over eight orders of magnitude (see inset) and a single copy of the target could be efficiently detected. The No Template Control (NTC) reaction generated no measurable fluorescence. Melt-curve analysis (below) demonstrates that no significant primer-dimers were produced because of the HotStart-IT binding protein (see NTC). | |
*As published in Applied Biosystem’s 2009 Molecular & Cell Biology Product Catalog (page 81).
