background: asthma is caused by the combination of different factors. current concepts of asthma pathogenesis emphasize on gene-environment interactions. mega-genome scanning projects revealed that different single nucleotide polymorphisms (snps) are related to asthma susceptibility. rs7216389-t is one of them that is related to childhood asthma and its effect on childhood asthma severity has b...

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 simple and rapid method to determine the G+C content of bacterial chromosomal DNA was developed. It involves determination of Tm by a Light Cycler and calculation of the G+C content by an empirical formula relating Tm to G+C content. Instead of a conventional thermal denaturation method, which monitors the increase of absorbance at 260 nm, thermal denaturation was monitored by the decrease of...

The CYP2C subfamily of the cytochrome P450 gene superfamily encodes heme-thiolate proteins that have a myriad of biologic functions. CYP2C proteins detoxify xenobiotics and metabolize endogenous lipids such as arachidonic acid to bioactive eicosanoids. We report new methods and results for the quantitative polymerase reaction (qPCR) analysis for the 15 members of the mouse Cyp2c subfamily (Cyp2...

Fluorescence resonance energy transfer (FRET) is a simple procedure for detecting specific DNA sequences, and is therefore used in many fields. However, the cost is relatively high, because FRET-based methods usually require fluorescent probes. We have designed a cost-effective way of using FRET, and developed a novel approach for the genotyping of single nucleotide polymorphisms (SNPs) and all...

Standardized procedures must be followed when characterizing, officially describing, and validly naming novel bacteria. For species descriptions, DNA-DNA hybridization still is needed for whole-genome comparisons between close relatives, but many established hybridization methods have drawbacks, such as requiring labeled or large amounts of DNA. We evaluated a new technique based on the spectro...

Next-generation sequencing (NGS) is becoming a standard for genetic analyses of clinical samples. DNAs retrieved from formalin-fixed, paraffin-embedded (FFPE) tissue specimens are commonly degraded, and specimens such as core biopsies are sometimes too small to obtain enough DNA for NGS applications. Thus, it is important to measure both the DNA quantity and quality accurately from clinical sam...

This work presents a method to visualize the degradation of exogenous DNA in living cells using a novel type of activatable fluorescence imaging probe. Deoxyribonuclease (DNase)-activatable fluorescence probes (DFProbes) are composed of double strands deoxyribonucleic acid (dsDNA) which is labeled with fluorophore (ROX or Cy3) and quencher on the end of one of its strands, and stained with SYBR...

This paper describes the use of real-time PCR for the confirmation of microarray data. Current publication guidelines require that all microarray results are confirmed by an independent gene expression profiling method. Real-time PCR is the method of choice for most researchers but it is not without drawbacks. The first step in confirming array results by real-time PCR is selection of gene-spec...