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KAPA SYBR® FAST qPCR Kits are designed for the highest performance in real-time PCR. Kits contain a novel DNA polymerase engineered through a process of molecular evolution - the result is a unique enzyme, specifically evolved for qPCR using SYBR® Green I dye chemistry.
The KAPA SYBR® DNA Polymerase was engineered to perform optimally in stringent real-time qPCR reaction conditions, exhibiting dramatic improvements to signal-to-noise ratio (fluorescence), cycle threshold (Ct), linearity, and sensitivity. The KAPA SYBR® DNA Polymerase and proprietary buffer system enhances the amplification efficiency of difficult templates, including both GC-rich and AT-rich templates.
"I lead genome-wide studies on RNA (gene expression and alternative splicing) and DNA (SNPs and copy number variation) and real-time PCR is a key player for validating our results. I switched to the KAPA SYBR® FAST qPCR Kit from Kapa Biosystems because I noticed a significant improvement in PCR reaction efficiency - the kit works very nicely for the vast majority of primers we design without optimization and for all our various applications. This universality makes our real-time PCR assays very fast to establish and our results more reliable."
- Dr. Philippe Demouging, Department of Molecular Psychology, University of Basel, Switzerland
Product ApplicationsKAPA SYBR® FAST qPCR Kits are ideally suited for: - Gene expression analysis
- Low copy gene detection
- Microarray validation
- Gene knockdown validation
- High-throughput qPCR
- ChIP
- Next-generation DNA sequencing library quantification
High performance gene expression analysisThe improved speed, processivity and robustness of KAPA SYBR® FAST qPCR Kits results in consistently high amplification efficiencies required for accurate relative quantification. To demonstrate the high performance of KAPA SYBR® FAST for gene expression analysis, the reaction efficiencies obtained for ten commonly used housekeeping genes in the human breast cancer cell line, MCF-7, were compared. The KAPA SYBR® FAST qPCR Kit achieved consistently high amplification efficiencies (95 – 104%) across all ten genes, despite differences in amplicon length. + No bias in amplification efficiency relative to GC content (44.2 – 62.5%) or amplicon length (86 – 249 bp) is observed with KAPA SYBR® FAST. Amplification efficiencies achieved for ten housekeeping genes with the KAPA SYBR® FAST Universal qPCR Kit, plotted against GC content (top) or amplicon length (bottom). The reaction efficiency achieved for each of the ten genes fell within the optimal range of 95 –105%, independent of the nature or length of the amplicon.
+ Typical results achieved with KAPA SYBR® FAST qPCR Kits in the amplification of housekeeping genes for gene expression analysis. Two housekeeping genes (ActB and HPRT1) were amplified from log-fold serial dilutions of MCF-7 cDNA (100 ng - 10 pg/reaction) using the KAPA SYBR® FAST Universal qPCR Kit (green) or the Invitrogen Express SYBR® GreenER™ Kit (blue). Linear amplification plots (top) demonstrate earlier Ct scores and greater baseline subtracted fluorescence for both the ActB and HPRT1 genes with the KAPA SYBR® FAST Kit. Calculated reaction efficiencies (bottom) confirmed that the consistently high performance required for accurate expression quantitation is achievable with the KAPA SYBR® FAST qPCR Kit (EActB = 100% and EHPRT1 = 104%). Efficiencies obtained with the Invitrogen Express SYBR® GreenER™ Kit were sub-optimal for both housekeeping genes (EActB= 107% and EHPRT1= 115%).
Superior signals, Ct values, and reaction efficienciesThe high performance of the KAPA SYBR® FAST qPCR kit can been seen when compared to other leading Fast qPCR Mater Mixes. KAPA SYBR® FAST consistently produces higher signal, earlier Cts, and 100% reaction efficiencies. + + + |
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