Efficient qPCR relies on good assay design. Since the invention of PCR, many parameters have been identified as important for assay quality, such as estimates of oligo temperature characteristics, GC content and folding properties.
Method outlines use of a hot start Taq for multiplex qPCR and provides guidance on how to optimize dNTPs, primer, probes and MgCL2 concentrations. By optimizing these parameters, the user can improve assay sensitivity and linear range of detection.
Our qPCR services, including primer and probe designs, assay protocol development, troubleshooting, and data analysis support, adhere to MIQE, which allows you to publish or bring your product to market faster and with confidence.
Examples of basic PCR/qPCR/dPCR protocols that can be used as the foundation for explorations into some of the concepts described in the theoretical chapters of this guide. Included are detailed protocols for assay quality control, in addition to more general
To achieve accurate and reliable results from your real-time quantitative PCR, choose from the broadest range of sequence–specific, fluorescent probes: LightCycler probes, Dual-labeled fluorogenic probes, Molecular Beacons, Scorpions™ probes. All of our fluorescent probes are deprotected, desalted and purified
The primer pairs have been designed to detect the most prevalent one for each eukaryotic gene based on a literature review. However, it is possible that the primer pairs may co-amplify other transcripts.
In recent years, array-based Comparative Genomic Hybridization (aCGH) has been refined to determine chromosomal changes at progressively higher resolutions. This evolving technology is, however, hampered by the large DNA input requirement—a minimum of 150,000 copies of a human genome, or
While many PCR assays are developed for research applications there are further considerations for those that are being developed to become diagnostic assays or to be performed in support of: Biologics License Application (BLA), New Drug Application (NDA), Premarket Approval
Probe based QPCR utilizes a fluorescent–labeled target-specific probe resulting in increased specificity and sensitivity. Additionally, a variety of fluorescent dyes are available so that multiple primers can be used to simultaneously amplify many sequences.
Developing a PCR or RT-PCR/RT-qPCR troubleshooting protocol so that data are reliable is essential. Potential sources of RT-PCR or PCR error and problems include operator error, the PCR master mix, and oligo design. This PCR troubleshooting guide outlines and details
Quantitative PCR (qPCR) provides information about gene expression, gene amplification or loss, and small alterations. qPCR is often used to investigate tumor biology and to discover the genetic and epigenetic causes of cancer