TaqMan, a variation of fluorescent PCR, is a powerful tool for gene expression and polymorphism studies. Here we describe the design and evaluation of 27 new TaqMan primer-probe sets for rat genes that play a key role in neural signaling. These newly designed and synthesized probes were tested and then used for quantification of RNA isolated from rat brain. The usual length of common TaqMan pro...

LEVEL: INTERMEDIATE R esidual host cell DNA removal in the processing of biological pharmaceuticals is an important metric for consistency of the purification process. There are also safety concerns with respect to the oncogenic potential of residual DNA from continuous cell lines. The acceptable residual amount of DNA is 10 ng/dose for parentally administrated drugs produced from continuous ce...

Applied Biosystems has designed and manufactured over 40,000 real-time PCR assays for measuring the expression of genes in humans, mice, and rats. TaqMan Gene Expression Assays each consist of amplification primers (forward and reverse) and a fluorescent labeled TaqMan probe, formulated into a single tube. One of the major user concerns regarding any real-time PCR-based assay product (primers a...

The aim of this study was to develop a rapid, sensitive and robust procedure for the qualitative detection of SARS coronavirus RNA. Three unique detection formats were developed for real-time RNA amplification assays: a post amplification detection step with a virus-specific internal capture probe based on Taqman (RT-PCR TaqMan assay), hybridization probe (RT-PCR hybridization probe assay) and ...

Since its first use, real-time quantitative PCR (qPCR) has evolved into a flexible, application-made method for the quantification and identification of nucleic acids (1-2). Depending on the application, most researchers choose between fluorescent nucleic acid binding dyes or labels that interact by fluorescence resonance energy transfer (FRET) as nucleic acid detection methods (2-3). Binding d...

Recent events illustrate the imperative to rapidly and accurately detect and identify pathogens during disease outbreaks, whether they are natural or engineered. Particularly for our primary goal of detecting bioterrorist releases, detection techniques must be both species-wide (capable of detecting all known strains of a given species) and species specific. Due to classification restrictions o...

The multiplexing capabilities with different fluorescent dyes are limited in real-time PCR instruments equipped with one excitation source. Considering this limitation, a design was developed to create a triple-labeled probe as an internal positive control (IPC) that utilizes a combination of the fluorescence resonance energy transfer (FRET) and TaqMan techniques. The IPC probe, labeled with FA...

The TaqMan® SNP genotyping technology utilizes the 5’ nuclease activity of Taq polymerase to generate a fluorescent signal during PCR. For each SNP, the assay uses two TaqMan probes that differ in sequence only at the SNP site, with one probe complementary to the wild-type allele and the other to the variant allele. The technique utilizes the FRET technology whereby a 5’ reporter dye and a 3’ q...

BACKGROUND The real-time polymerase chain reaction is currently the method of choice for quantifying nucleic acids in different DNA based quantification applications. It is widely used also for detecting and quantifying genetically modified components in food and feed, predominantly employing TaqMan and SYBR Green real-time PCR chemistries. In our study four alternative chemistries: Lux, Plexor...