qRT-PCR: a method and its difficulties

Development and applications of a real-time quantitative RT-PCR method (QRT-PCR) for BRCA1 mRNA.

OBJECTIVES To develop a real-time quantitative RT-PCR method for BRCA1 mRNA and then use it for the study of BRCA1 gene expression in human MCF-7 breast cancer cells after their exposure to antineoplastic agents and gamma irradiation. DESIGN AND METHODS The developed QRT-PCR method is based on the real-time monitoring of a fluorescein-labeled TaqMan probe, specific for BRCA1 mRNA, during PCR ...

Real-time reverse transcription PCR (qRT-PCR) and its potential use in clinical diagnosis.

qRT-PCR (real-time reverse transcription-PCR) has become the benchmark for the detection and quantification of RNA targets and is being utilized increasingly in novel clinical diagnostic assays. Quantitative results obtained by this technology are not only more informative than qualitative data, but simplify assay standardization and quality management. qRT-PCR assays are most established for t...

Evaluation of bacterial internalization using qRT-PCR.

1Bacterial resistance to antibiotics currently represents a topic of high interest. Unfortunately, studies addressing their adaptation and multidrug tolerance latency are not sufficient for a comprehensive description of the pathogen-host molecular interaction mechanisms (interactome) and the bacterial internalization into human cells, which leads to modified human genes. Bacterial persisters a...

Detection of Langat virus by TaqMan real-time one-step qRT-PCR method

Langat virus (LGTV), one of the members of the tick-borne encephalitis virus (TBEV) complex, was firstly isolated from Ixodes granulatus ticks in Malaysia. However, the prevalence of LGTV in ticks in the region remains unknown. Surveillance for LGTV is therefore important and thus a tool for specific detection of LGTV is needed. In the present study, we developed a real-time quantitative revers...

Quantitative Real-time Polymerase Chain Reaction (qRT-PCR). Fatemeh Mehrpooyan*