Following the initial report of the use of SYBR Green I for real-time polymerase chain reaction (PCR) in 1997, little attention has been given to the development of alternative intercalating dyes for this application. This is surprising considering the reported limitations of SYBR Green I, which include limited dye stability, dye-dependent PCR inhibition, and selective detection of amplicons du...

A real-time multiplex PCR procedure with melting curve analysis, using the green fluorescence dye SYBR Green I, was developed for rapid and reliable identification of Plum pox virus (PPV) isolates of strains D and M. Members of the different strains were identified by their distinctive melting temperatures (T(m)s); 84.3-84.43 degrees C for D isolates, and 85.34-86.11 degrees C for M isolates. T...

Microbial cell counting provides essential information for the study of abundance profiles and biogeochemical interactions with surrounding environments. However, it often requires labor-intensive time-consuming processes, particularly subseafloor sediment samples, in which non-cell particles are abundant. We developed a rapid straightforward method staining microbial intracellular DNA by SYBR ...

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...

Real-time quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) is the method of choice for rapid and reproducible measurements of cytokine or growth factor expression in small samples. Fluorescence detection methods for monitoring real-time PCR include fluorogenic probes labelled with reporter and quencher dyes, such as Taqman probes or Molecular Beacons and the dsDNA-binding d...

background: breast cancer is the most common cancer in women. non-coding rnasespecially mirnas have important regulatory roles in cancer. mirnas are 21-24 nucleotides which have different levels of expression between tumors and normal tissues. in this study, we have analyzed expression level of mir-520d in three different groups of breast cancer. methods: fifty nine samples were divided into di...

Since its first use, real-time quantitative PCR (qPCR) has evolved into a flexible, application-made method for the quantification and identification of nucleic acids (1-2). Depending on the application, most researchers choose between fluorescent nucleic acid binding dyes or labels that interact by fluorescence resonance energy transfer (FRET) as nucleic acid detection methods (2-3). Binding d...

A comparative study was conducted of a novel real-time quantitative PCR test (LightCycler System) with FastStart DNA Master(PLUS) SYBR Green I dye to detect DNA of human herpes virus 6 (HHV-6). Results were compared with those of a real-time quantitative PCR with hybridization probe (HP) formats using the fluorescence resonance energy transfer method, and with those of a single qualitative PCR ...

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...