Prime time for real-time PCR

  title={Prime time for real-time PCR},
  author={Laura P. Bonetta},
  journal={Nature Methods},
  • L. Bonetta
  • Published 1 April 2005
  • Biology
  • Nature Methods
Real-time PCR is the favored method for measuring gene expression. Researchers benefit from a vast and growing choice of reagents and instruments for their experiments. Laura Bonetta reports. 

Gene expression: one size does not fit all

Although in recent years the field has been flooded with microarray data, a multitude of non–array-based methods for studying the expression of genes are on the market.

Ligation-Enabled Fluorescence-Coding PCR for High-Dimensional Fluorescence-Based Nucleic Acid Detection.

The LiNC PCR technique is demonstrated by implementing a two-color-based assay for detection of 10 ovarian cancer epigenetic biomarkers at analytical sensitivities as low as 60 template molecules with no detectable target cross-talk.

PCR-Stop analysis as a new tool for qPCR assay validation

The benefits of PCR-Stop analysis reveal quantitative and qualitative resolution of both assays, the limits of one of those assays and thus avoiding misinterpretations in qPCR analysis, and data displayed that a well performing assay starts indeed with its average efficiency.

A Fast-and-Robust Profiler for Improving Polymerase Chain Reaction Diagnostics

A profiling method is proposed that permits the non-linear maximization of amplicon resolution by eliminating the necessity for direct error estimation and is expected to have extensive applications in genetics and biotechnology where there is a demand for cheap, expedient, and robust information.

Real-time PCR assay for the discrimination and quantification of wheat and barley strains of Wheat dwarf virus

A rapid method for detection, discrimination and quantification of wheat and barley strains of wheat dwarf virus (WDV) was successfully developed and both TaqMan® and SYBR® Green technologies provided accurate and reliable methods.

Fluorescence Polarization Based Nucleic Acid Testing for Rapid and Cost-Effective Diagnosis of Infectious Disease.

A new nucleic acid detection method was developed for a rapid and cost-effective diagnosis of infectious disease and was used to detect and sub-type Salmonella bacteria with sensitivities down to a single bacterium in less than three hours.

Detection and Serotype-Specific Differentiation of Vesicular Stomatitis Virus Using a Multiplex, Real-Time, Reverse Transcription-Polymerase Chain Reaction Assay

  • K. HoleA. ClavijoLuis A Pineda
  • Medicine, Biology
    Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc
  • 2006
The real-time RT-PCR assay was documented to be sensitive and specific for the detection ofVSV-NJ and VSV-IN (1–3) strains from field samples in a single reaction, thereby supporting use of this assay in the differential diagnosis of vesicular virus diseases in cattle.

The Qiagen Investigator® Quantiplex HYres as an alternative kit for DNA quantification.



RNAi: Silencing never sounded better

The ability to trigger RNA interference in mammalian cells provides unprecedented opportunities for probing the functions of genes. Many products and resources are there to help. Laura Bonetta

Linear-After-The-Exponential (LATE)–PCR: An advanced method of asymmetric PCR and its uses in quantitative real-time analysis

Conventional asymmetric PCR is inefficient and difficult to optimize because limiting the concentration of one primer lowers its melting temperature below the reaction annealing temperature.

Detection of PCR products using self-probing amplicons and fluorescence

This work describes a new technology that is simple to use, gives highly specific information, and avoids the major difficulties of the alternative methods of molecular diagnostics.

Wavelength-shifting molecular beacons

Wavelength-shifting molecular beacons are substantially brighter than conventional molecularBeacons that contain a fluorophore that cannot efficiently absorb energy from the available monochromatic light source, and their use improves and simplifies multiplex genetic analyses.

Real-time measurement of in vitro transcription.

A simple method to measure RNA synthesis in real time was developed, and it was found that molecular beacons synthesized from natural deoxyribonucleotides were not suitable, because they are copied by RNA polymerases, generating complementary product strands that bind to the Molecular beacons, causing a conformational change that results in unwanted fluorescence.

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Laura Bonetta is a freelance writer based in the Washington, DC area. (

  • Laura Bonetta is a freelance writer based in the Washington, DC area. (