Real-Time PCR

Since its introduction in 1993, real-time PCR – also known as quantitative PCR (qPCR) – has become one of the fastest growing PCR applications. Based on the principles of conventional or end-point PCR, qPCR utilizes fluorophores to continuously collect data in the form of fluorescent signals from one or more primer-template interactions over a range of PCR cycles. Non-sequence specific assays employ intercalating fluorescent dyes such SYBR® Green I to detect the accumulation of double-stranded DNA PCR product, whereas sequence specific assays are based on the use of fluorescently labeled probes (e.g. TaqMan® probes used in 5’-nuclease assays, FRET probes or molecular beacons). Fluorescent signals from each reaction are converted into a numerical value for each sample. qPCR has become the most sensitive and specific quantitative method for quantitative gene expression, genotyping, SNP analysis, pathogen detection and drug target validation. In addition to the extremely broad dynamic range and high specificity, the closed-tube qPCR format is ideally suited to high sample throughput and limits the risk of sample contamination associated with post-amplification handing.

Kapa Biosystems offers a suite of qPCR reagents, both for sequence non-specific and sequence specific assays:

KAPA SYBR® FAST qPCR Kits are designed for high performance SYBR® Green I-based real-time PCR. The Master Mixes supplied in these kits contain a novel DNA polymerase, specifically engineered (using molecular evolution) for fast, high efficiency qPCR in the presence of SYBR® Green I. The major benefits of KAPA SYBR® FAST qPCR Kits are:

  • Superior sensitivity, linearity and reaction efficiency across a wide range of template types and assays.
  • Superior signal intensities, due to the inclusion of higher concentrations of SYBR® Green I dye.
  • Reduced cycling times, which reduce overall turnaround times.
  • Increased flexibility in primer design due to the ability to amplify longer targets.

KAPA PROBE FAST qPCR Kits are designed for probe-based qPCR. These kits have been optimized for versatility, reproducibility and speed. Applications include genotyping, gene expression analysis, pathogen detection and drug target validation. The major benefits of KAPA PROBE FAST qPCR Kits are:

  • Compatibility with all probe-based applications and instruments.
  • Fast, reproducible and precise quantification.
  • Discrete clusters in SNP genotyping assays.
  • Suitability for multiplex qPCR.
  • Broad dynamic range.
  • Highly stable master mixes for high throughput workflows.

Fast Real-Time PCR

KAPA SYBR® FAST and KAPA PROBE FAST qPCR Kits have been optimized for high-speed, real-time PCR without loss in performance.