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.
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
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
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
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
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.
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.
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
The entire PCR workflow is vulnerable to factors which introduce variability. Many of the variable components are unavoidable, such as the source of the sample or the requirement for a reverse transcription step. Assay design is also highly variable and