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Viability PCR
Viability PCR, also named v-PCR or vPCR, is an evolution of Polymerase chain reaction, PCR. Through the use of a simple pre-treatment of the sample by the means of specific intercalating photo-reactive reagents it's possible to neutralize the DNA of dead cells. As a result, only DNA from live cells will be detected by PCR. This approach expands a lot the analytical scope of PCR procedures. The capability to detect only living cells become very important, because in key applications is more important to know the amount of live cells, than the total cell level. Examples of this are: food and water quality control, infectious diseases diagnostic, veterinary applications, ecological dynamics... The first referenced work about this analytical approach was in 2003, Norwegian researchers suggest the use of Ethidium Monoazide, an azide form of Ethidium Bromide, which was used in other analytical fields as Flow cytometry, Flow Cytometry as a candidate for viability PCR. However, the main ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Polymerase Chain Reaction
The polymerase chain reaction (PCR) is a method widely used to rapidly make millions to billions of copies (complete or partial) of a specific DNA sample, allowing scientists to take a very small sample of DNA and amplify it (or a part of it) to a large enough amount to study in detail. PCR was invented in 1983 by the American biochemist Kary Mullis at Cetus Corporation; Mullis and biochemist Michael Smith (chemist), Michael Smith, who had developed other essential ways of manipulating DNA, were jointly awarded the Nobel Prize in Chemistry in 1993. PCR is fundamental to many of the procedures used in genetic testing and research, including analysis of Ancient DNA, ancient samples of DNA and identification of infectious agents. Using PCR, copies of very small amounts of DNA sequences are exponentially amplified in a series of cycles of temperature changes. PCR is now a common and often indispensable technique used in medical laboratory research for a broad variety of applications ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Ethidium Bromide
Ethidium bromide (or homidium bromide, chloride salt homidium chloride) is an intercalating agent commonly used as a fluorescent tag (nucleic acid stain) in molecular biology laboratories for techniques such as agarose gel electrophoresis. It is commonly abbreviated as EtBr, which is also an abbreviation for bromoethane. To avoid confusion, some laboratories have used the abbreviation EthBr for this salt. When exposed to ultraviolet light, it will fluoresce with an orange colour, intensifying almost 20-fold after binding to DNA. Under the name homidium, it has been commonly used since the 1950s in veterinary medicine to treat trypanosomiasis in cattle. The high incidence of antimicrobial resistance makes this treatment impractical in some areas, where the related isometamidium chloride is used instead. Despite its reputation as a mutagen, it is relatively safe to handle. Structure, chemistry, and fluorescence As with most fluorescent compounds, ethidium bromide is aromatic. ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Flow Cytometry
Flow cytometry (FC) is a technique used to detect and measure physical and chemical characteristics of a population of cells or particles. In this process, a sample containing cells or particles is suspended in a fluid and injected into the flow cytometer instrument. The sample is focused to ideally flow one cell at a time through a laser beam, where the light scattered is characteristic to the cells and their components. Cells are often labeled with fluorescent markers so light is absorbed and then emitted in a band of wavelengths. Tens of thousands of cells can be quickly examined and the data gathered are processed by a computer. Flow cytometry is routinely used in basic research, clinical practice, and clinical trials. Uses for flow cytometry include: * Cell counting * Cell sorting * Determining cell characteristics and function * Detecting microorganisms * Biomarker detection * Protein engineering detection * Diagnosis of health disorders such as blood cancers * Measuring ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Propidium Monoazide
Propidium monoazide (PMA) is a photoreactive DNA-binding dye that preferentially binds to dsDNA. It is used to detect viable microorganisms by qPCR. Visible light (high power halogen lamps or specific LED devices) induces a photoreaction of the chemical that will lead to a covalent bond with PMA and the dsDNA. The mechanism of DNA modification by PMA can be seen in this protocol. This process renders the DNA insoluble and results in its loss during subsequent genomic DNA extraction. Theoretically, dead microorganisms lose their capability to maintain their membranes intact, which leaves the "naked" DNA in the cytosol ready to react with PMA. DNA of living organisms are not exposed to the PMA, as they have an intact cell membrane. After treatment with the chemical, only the DNA from living bacteria is usable in qPCR, allowing to obtain only the amplified DNA of living organisms. This is helpful in determining which pathogens are active in specific samples. The main use of PMA is in ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Dyes
A dye is a colored substance that chemically bonds to the substrate to which it is being applied. This distinguishes dyes from pigments which do not chemically bind to the material they color. Dye is generally applied in an aqueous solution and may require a mordant to improve the fastness of the dye on the fiber. There are two broad categories of dyes: natural and synthetic; Natural dyes are dyes extracted from plants, Insects, or minerals. The majority of natural dyes are vegetable dyes derived from plant sources such as roots, berries, bark, leaves, and wood, as well as other biological sources like fungi. Synthetic dyes are also referred to as "coal tar dyes" because they are derived from substances that, until recently, could only be extracted from coal tar. A synthetic dye consists of a chromophore and an auxochrome added to a benzene derivative. Both dyes and pigments are colored, because they absorb only some wavelengths of visible light. Dyes are usually soluble i ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Molecular Biology
Molecular biology is the branch of biology that seeks to understand the molecular basis of biological activity in and between cells, including biomolecular synthesis, modification, mechanisms, and interactions. The study of chemical and physical structure of biological macromolecules is known as molecular biology. Molecular biology was first described as an approach focused on the underpinnings of biological phenomena - uncovering the structures of biological molecules as well as their interactions, and how these interactions explain observations of classical biology. In 1945 the term molecular biology was used by physicist William Astbury. In 1953 Francis Crick, James Watson, Rosalind Franklin, and colleagues, working at Medical Research Council unit, Cavendish laboratory, Cambridge (now the MRC Laboratory of Molecular Biology), made a double helix model of DNA which changed the entire research scenario. They proposed the DNA structure based on previous research done by Ro ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
