For the rapid identification and detection of Vibrio anguillarum, we have PCR amplification technique targeting gyrB region has been evaluated. We have designed twosets of PCR primers for the specific amplification of gyrB in V. anguillarum using single and nested PCR. PCRs specificity was demonstrated by successful amplicon from V. anguillarum DNA. The detection limit of the single PCRs and th...

This study developed a new multiplex polymerase chain reaction (m-PCR) for rapidly detecting clinically essential strains of V. parahaemolyticus. enables the detection total and potentially virulent strains. The m-PCR was by targeting species-specific transcriptional regulator toxR gene, sequences an outer membrane protein hypothetical encoded omp htp, respectively. htp were discovered original...

We present a high throughput microfluidic device for continuous-flow polymerase chain reaction (PCR) in water-in-oil droplets of nanoliter volumes. The circular design of this device allows droplets to pass through alternating temperature zones and complete 34 cycles of PCR in only 17 min, avoiding temperature cycling of the entire device. The temperatures for the applied two-temperature PCR pr...

Real-time reverse transcription-polymerase chain reaction (RT-PCR) is currently considered the most sensitive method to study low abundance gene expression. Since comparison of gene expression levels in various tissues is often the purpose of an experiment, we studied a tissue-linked effect on nucleic acid amplification. Based on the raw data generated by a LightCycler instrument, we propose a ...

Polymerase Chain Reaction (PCR) is a major DNA amplification technology from molecular biology. The quantitative analysis of PCR aims at determining the initial amount of the DNA molecules from the observation of typically several PCR amplifications curves. The mainstream observation scheme of the DNA amplification during PCR involves fluorescence intensity measurements. Under the classical ass...

Polymerase chain reaction (PCR) is a major DNA amplification technology from molecular biology. The quantitative analysis of PCR aims at determining the initial amount of the DNA molecules from the observation of typically several PCR amplifications curves. The mainstream observation scheme of the DNA amplification during PCR involves fluorescence intensity measurements. Under the classical ass...

Current quantitative PCR protocols are only indicative of the relative quantity of a target sequence to a standard since no means of estimating the amplification rate is available as yet. The variability of PCR performed on isolated cells has already been reported by several authors, but it could not be extensively studied due to lack of a system for doing kinetic data acquisition and of statis...

Nucleic acid amplification can provide sensitive and specific molecular detection and quantification. Since its introduction PCR has been the dominant amplification technology; its widespread use has stimulated improvements in enzymes, instrumentation, and analytical methods. Initial use of PCR was largely qualitative: positive target detection, or inference of target absence above a detection ...

Methylation analysis of individual cytosines in genomic DNA can be determined quantitatively by bisulphite treatment and PCR amplification of the target DNA sequence, followed by restriction enzyme digestion or sequencing. Methylated and unmethylated molecules, however, have different sequences after bisulphite conversion. For some sequences this can result in bias during the PCR amplification ...