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RT-PCR Test
Reverse transcription polymerase chain reaction (RT-PCR) is a laboratory technique combining reverse transcription of RNA into DNA (in this context called complementary DNA or cDNA) and amplification of specific DNA targets using polymerase chain reaction (PCR). It is primarily used to measure the amount of a specific RNA. This is achieved by monitoring the amplification reaction using fluorescence, a technique called real-time PCR or quantitative PCR (qPCR). Combined RT-PCR and qPCR are routinely used for analysis of gene expression and quantification of viral RNA in research and clinical settings. The close association between RT-PCR and qPCR has led to metonymic use of the term qPCR to mean RT-PCR. Such use may be confusing, as RT-PCR can be used without qPCR, for example to enable molecular cloning, sequencing or simple detection of RNA. Conversely, qPCR may be used without RT-PCR, for example to quantify the copy number of a specific piece of DNA. Nomenclature The combined ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Virus
A virus is a submicroscopic infectious agent that replicates only inside the living cells of an organism. Viruses infect all life forms, from animals and plants to microorganisms, including bacteria and archaea. Since Dmitri Ivanovsky's 1892 article describing a non-bacterial pathogen infecting tobacco plants and the discovery of the tobacco mosaic virus by Martinus Beijerinck in 1898,Dimmock p. 4 more than 9,000 virus species have been described in detail of the millions of types of viruses in the environment. Viruses are found in almost every ecosystem on Earth and are the most numerous type of biological entity. The study of viruses is known as virology, a subspeciality of microbiology. When infected, a host cell is often forced to rapidly produce thousands of copies of the original virus. When not inside an infected cell or in the process of infecting a cell, viruses exist in the form of independent particles, or ''virions'', consisting of (i) the genetic material, i. ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Scintillation Counting
Liquid scintillation counting is the measurement of radioactive activity of a sample material which uses the technique of mixing the active material with a liquid scintillator (e.g. zinc sulfide), and counting the resultant photon emissions. The purpose is to allow more efficient counting due to the intimate contact of the activity with the scintillator. It is generally used for alpha particle or beta particle detection. Technique Samples are dissolved or suspended in a "cocktail" containing a solvent (historically aromatic organics such as xylene or toluene, but more recently less hazardous solvents are used), typically some form of a surfactant, and "fluors" or scintillators which produce the light measured by the detector. Scintillators can be divided into primary and secondary phosphors, differing in their luminescence properties. Beta particles emitted from the isotopic sample transfer energy to the solvent molecules: the π cloud of the aromatic ring absorbs the energy of t ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Phosphorimager
Photostimulated luminescence (PSL) is the release of stored energy within a phosphor by stimulation with visible light, to produce a luminescent signal. X-rays may induce such an energy storage. A plate based on this mechanism is called a photostimulable phosphor (PSP) plate and is one type of X-ray detector used in projectional radiography. Creating an image requires illuminating the plate twice: the first exposure, to the radiation of interest, "writes" the image, and a later, second illumination (typically by a visible-wavelength laser) "reads" the image. The device to read such a plate is known as a phosphorimager (occasionally spelled phosphoimager, perhaps reflecting its common application in molecular biology for detecting radiolabeled phosphorylated proteins and nucleic acids). Projectional radiography using a photostimulable phosphor plate as an X-ray detector can be called "phosphor plate radiography" or "computed radiography" (not to be confused with computed tomography w ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Phosphorus-32
Phosphorus-32 (32P) is a radioactive isotope of phosphorus. The nucleus of phosphorus-32 contains 15 protons and 17 neutrons, one more neutron than the most common isotope of phosphorus, phosphorus-31. Phosphorus-32 only exists in small quantities on Earth as it has a short half-life of 14 days and so decays rapidly. Phosphorus is found in many organic molecules and so phosphorus-32 has many applications in medicine, biochemistry, and molecular biology where it can be used to trace phosphorylated molecules (for example, in elucidating metabolic pathways) and radioactively label DNA. Decay Phosphorus-32 has a short half-life of 14.268 days and decays into sulfur-32 by beta decay as shown in this nuclear equation: : 1.709 MeV of energy is released during the decay. The kinetic energy of the electron varies with an average of approximately 0.5 MeV and the remainder of the energy is carried by the nearly undetectable electron antineutrino. In comparison to other beta radiation-emi ... [...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] |
Melting Point
The melting point (or, rarely, liquefaction point) of a substance is the temperature at which it changes state from solid to liquid. At the melting point the solid and liquid phase exist in equilibrium. The melting point of a substance depends on pressure and is usually specified at a standard pressure such as 1 atmosphere or 100 kPa. When considered as the temperature of the reverse change from liquid to solid, it is referred to as the freezing point or crystallization point. Because of the ability of substances to supercool, the freezing point can easily appear to be below its actual value. When the "characteristic freezing point" of a substance is determined, in fact, the actual methodology is almost always "the principle of observing the disappearance rather than the formation of ice, that is, the melting point." Examples For most substances, melting and freezing points are approximately equal. For example, the melting point ''and'' freezing point of mercury is . How ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Primer-dimer
A primer dimer (PD) is a potential by-product in the polymerase chain reaction (PCR), a common biotechnological method. As its name implies, a PD consists of two primer molecules that have attached ( hybridized) to each other because of strings of complementary bases in the primers. As a result, the DNA polymerase amplifies the PD, leading to competition for PCR reagents, thus potentially inhibiting amplification of the DNA sequence targeted for PCR amplification. In quantitative PCR, PDs may interfere with accurate quantification. Mechanism of formation A primer dimer is formed and amplified in three steps. In the first step, two primers anneal at their respective 3' ends (step I in the figure). If this construct is stable enough, the DNA polymerase will bind and extend the primers according to the complementary sequence (step II in the figure). An important factor contributing to the stability of the construct in step I is a high GC-content at the 3' ends and length of the overl ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
SYBR Green
SYBR Green I (SG) is an asymmetrical cyanine dye used as a nucleic acid stain in molecular biology. The SYBR family of dyes is produced by Molecular Probes Inc., now owned by Thermo Fisher Scientific. SYBR Green I binds to DNA. The resulting DNA-dye-complex best absorbs 497 nanometer blue light (λmax = 497 nm) and emits green light (λmax = 520 nm). The stain preferentially binds to double-stranded DNA, but will stain single-stranded (ss) DNA with lower performance. SYBR Green can also stain RNA with a lower performance than ssDNA. Uses SYBR Green finds usage in several areas of biochemistry and molecular biology. It is used as a dye for the quantification of double stranded DNA in some methods of quantitative PCR. It is also used to visualise DNA in gel electrophoresis. Higher concentrations of SYBR Green can be used to stain agarose gels in order to visualise the DNA present. In addition to labelling pure nucleic acids, SYBR Green can also be used for labelling o ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Serum (blood)
Serum () is the fluid and solute component of blood which does not play a role in Coagulation, clotting. It may be defined as blood plasma without the clotting factors, or as blood with all cells and clotting factors removed. Serum includes all proteins not used in Coagulation, blood clotting; all electrolytes, antibodies, antigens, hormones; and any exogenous substances (e.g., drugs or microorganisms). Serum does not contain white blood cells (leukocytes), red blood cells (erythrocytes), platelets, or clotting factors. The study of serum is serology. Serum is used in numerous diagnostic tests as well as blood typing. Measuring the concentration of various molecules can be useful for many applications, such as determining the therapeutic index of a drug candidate in a clinical trial. To obtain serum, a blood sample is allowed to clot (coagulation). The sample is then centrifuged to remove the clot and blood cells, and the resulting liquid wikt:supernatant, supernatant is serum. ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Whole Blood
Whole blood (WB) is human blood from a standard blood donation. It is used in the treatment of massive bleeding, in exchange transfusion, and when people donate blood to themselves. One unit of whole blood (~517 mls) brings up hemoglobin levels by about 10 g/L. Cross matching is typically done before the blood is given. It is given by injection into a vein. Side effects include red blood cell breakdown, high blood potassium, infection, volume overload, lung injury, and allergic reactions such as anaphylaxis. Whole blood is made up of red blood cells, white blood cells, platelets, and blood plasma. It is best within a day of collection; however, can be used for up to three weeks. The blood is typically combined with an anticoagulant and preservative during the collection process. The first transfusion of whole blood was in 1818; however, common use did not begin until the First and Second World Wars. It is on the World Health Organization's List of Essential Medicines. ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
MRNA
In molecular biology, messenger ribonucleic acid (mRNA) is a single-stranded molecule of RNA that corresponds to the genetic sequence of a gene, and is read by a ribosome in the process of Protein biosynthesis, synthesizing a protein. mRNA is created during the process of Transcription (biology), transcription, where an enzyme (RNA polymerase) converts the gene into primary transcript mRNA (also known as pre-mRNA). This pre-mRNA usually still contains introns, regions that will not go on to code for the final amino acid sequence. These are removed in the process of RNA splicing, leaving only exons, regions that will encode the protein. This exon sequence constitutes mature mRNA. Mature mRNA is then read by the ribosome, and, utilising amino acids carried by transfer RNA (tRNA), the ribosome creates the protein. This process is known as Translation (biology), translation. All of these processes form part of the central dogma of molecular biology, which describes the flow of genet ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
