The SYBR green I (SG) dye-based fluorescence assay for screening antimalarial compounds is based on direct quantitation of parasite DNA. We show that DNA-interacting cationic cell-penetrating peptides (CPPs) and intercalating agents compete with SG dye to bind to DNA. Therefore, readouts of this assay, unlike those of the [(3)H]hypoxanthine incorporation assay, for the antimalarial activity of ...

Lyme disease caused by Borrelia burgdorferi is the most common tick-borne disease in the US and Europe. Unlike most bacteria, measurements of growth and viability of B. burgdorferi are challenging. The current B. burgdorferi viability assays based on microscopic counting and PCR are cumbersome and tedious and cannot be used in a high throughput format. Here, we evaluated several commonly used v...

Background: Watermelon silver mottle virus (WSMoV), which belongs to the genus Tospovirus, causes significant loss in Cucurbitaceae plants. Objectives: Development of a highly sensitive and reliable detection method for WSMoV. Materials and Methods: Recombinant plasmids for targeting the sequence of nucleocapsid protein gene of WSMoV were constructed. SYBR Green I real-time PCR was established...

EvaGreen is a new DNA intercalating dye successfully used in quantitative real-time PCR. In the present work, we firstly apply EvaGreen to the analysis of dsDNA by CE with LIF detection. Comparisons of EvaGreen dye with the commonly used dyes SYBR Green I and SYBR Gold were preformed in dsDNA analysis by CE. The linear range of dsDNA using EvaGreen was slightly wider than that using SYBR Gold a...

Flow cytometry can rapidly quantitate bacteria and discriminate them from debris. Researchers at the Swiss Federal Institute of Aquatic Science and Technology (Eawag, Switzerland) have developed a standard flow cytometric staining protocol and a corresponding BD AccuriTM C6 software analysis template to discriminate bacteria from debris in drinking water samples. The template employs a single, ...

background: watermelon silver mottle virus (wsmov), which belongs to the genus tospovirus, causes significant loss in cucurbitaceae plants. objectives: development of a highly sensitive and reliable detection method for wsmov.materials and methods: recombinant plasmids for targeting the sequence of nucleocapsid protein gene of wsmov were constructed. sybr green i real-time pcr was established a...

Background: The outbreak of novel influenza A H1N1-2009 virus (pH1N1) and its rapid spread worldwide raised serious concern about pandemic preparedness. Objectives: The present study was designed to evaluate the sensitivity and specificity of two chemistries of real-time RT-PCR (with the use of fluorescent SYBR Green I dye and specific TaqMan probe) for detection of the matrix and hemagglutinin...

Real-time PCR is an accurate method that can be used for the quantification of specific DNA molecules. Here we provide a protocol for SYBR Green I in real-time PCR applications using plastic reaction tubes. We report that SYBR Green I is alkali labile and once degraded inhibits the PCR. In our optimized protocol, diluted aliquots of SYBR Green I remain stable for at least two weeks. We also eva...

A label-free and sensitive fluorescence method for recognition of sequence-specific DNA using DNA-intercalating dye and metal-organic frameworks (MOFs) is developed. Here, MIL-101 (Cr3F(H2O)2O[(O2C)-C6H4-(CO2)]3·nH2O) is introduced as a quenching platform to decrease the high background fluorescence of SYBR Green I (SG)/probe DNA complex. Mechanism investigations show that MIL-101 can strongly ...