Method To Detect Only Live Bacteria during PCR Amplification

Method To Detect Only Live Bacteria during PCR Amplification.

Ethidium monoazide (EMA) is a DNA cross-linking agent and eukaryotic topoisomerase II poison. We previously reported that the treatment of EMA with visible light irradiation (EMA + Light) directly cleaved chromosomal DNA of Escherichia coli (T. Soejima, K. Iida, T. Qin, H. Taniai, M. Seki, A. Takade, and S. Yoshida, Microbiol. Immunol. 51:763-775, 2007). Herein, we report that EMA + Light rando...

Errors induced during PCR amplification

The Polymerase Chain Reaction (PCR) is one of the most widely used techniques in modern molecular biology to amplify a single or few copies of a piece of DNA sequences. The PCR products are used as inputs of many other applications such as diagnosis of diseases. Therefore the accuracy of the further analysis depends on the quality of the PCR outputs. In order to study the amplification process ...

TaqMan real-time PCR: a reliable method to detect meat species

The aim of this study was to detect and identify meat species (beef, lamb, pork, chicken, donkey, and horse) in Kebab as a Halal meat-based food sold in on-road-restaurants of North Khorasan province (Iran) using TaqMan real-time polymerase chain reaction (PCR). Raw Kebab samples (150 samples) were obtained from on-road restaurants. The samples were prepared, DNA was extracted and TaqMan real-t...

Failure of a PCR screening method to detect MRSA.

NcGRA2-RT-PCR to Detect Live Versus Dead Parasites in Neospora caninum-Infected Mice

In the present work, we optimized a recently established NcGRA2-RT-PCR based on RNA to detect live Neospora caninum parasites in tissue, and compared the results with the conventional inoculation of diagnostic specimen onto cell culture. C57BL/6 mice were experimentally infected with Nc-1 tachyzoites and subsequently euthanized 6 or 12 days post infection (dpi). Selected organs were used to sea...