Buoyant Density Ultracentrifugation
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Buoyant Density Ultracentrifugation
Buoyant density centrifugation (also isopycnic centrifugation or equilibrium density-gradient centrifugation) uses the concept of buoyancy to separate molecules in solution by their differences in density. Implementation Historically a caesium chloride (CsCl) solution was often used, but more commonly used density gradients are sucrose or Percoll. The sample is put on top of the solution, and then the tube is spun at a very high speed for an extended time, at times lasting days. The CsCl molecules become densely packed toward the bottom, so even layers of different densities form. Since the original solution was approximately the same density, they go to a level where their density and the CsCl density are the same, to which they form a sharp, distinctive band. Isotope separation This method very sharply separates molecules, and is so sharp that it can even separate different molecular isotopes from one another. DNA separation Buoyant density of majority of DNA is 1.7g/cm3 whi ...
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Canines Parvovirus
Canine may refer to: Zoology and anatomy * a dog-like Canid animal in the subfamily Caninae ** ''Canis'', a genus including dogs, wolves, coyotes, and jackals ** Dog, the domestic dog * Canine tooth, in mammalian oral anatomy People with the surname * Henry Canine (), American football coach * Ralph Canine (1895–1969), founding director of the United States National Security Agency Other uses * Canine, a fictional dog in the ''Glenn Martin, DDS'' animated television series * Canine Hills, Antarctic landform in the Bowers Mountains, Victoria Land See also * K9 (other) * Kanine (other) * Canina (other) * Cani (other) * List of canids Canidae is a family of mammals in the order Carnivora, which includes domestic dogs, wolves, coyotes, foxes, jackals, dingoes, and many other extant and extinct dog-like mammals. A member of this family is called a canid; all extant spec ...
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Buoyancy
Buoyancy (), or upthrust, is an upward force exerted by a fluid that opposes the weight of a partially or fully immersed object. In a column of fluid, pressure increases with depth as a result of the weight of the overlying fluid. Thus the pressure at the bottom of a column of fluid is greater than at the top of the column. Similarly, the pressure at the bottom of an object submerged in a fluid is greater than at the top of the object. The pressure difference results in a net upward force on the object. The magnitude of the force is proportional to the pressure difference, and (as explained by Archimedes' principle) is equivalent to the weight of the fluid that would otherwise occupy the submerged volume of the object, i.e. the displaced fluid. For this reason, an object whose average density is greater than that of the fluid in which it is submerged tends to sink. If the object is less dense than the liquid, the force can keep the object afloat. This can occur only in a no ...
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Caesium Chloride
Caesium chloride or cesium chloride is the inorganic compound with the formula Cs Cl. This colorless salt is an important source of caesium ions in a variety of niche applications. Its crystal structure forms a major structural type where each caesium ion is coordinated by 8 chloride ions. Caesium chloride dissolves in water. CsCl changes to NaCl structure on heating. Caesium chloride occurs naturally as impurities in carnallite (up to 0.002%), sylvite and kainite. Less than 20 tonnes of CsCl is produced annually worldwide, mostly from a caesium-bearing mineral pollucite. Caesium chloride is widely used medicine structure in isopycnic centrifugation for separating various types of DNA. It is a reagent in analytical chemistry, where it is used to identify ions by the color and morphology of the precipitate. When enriched in radioisotopes, such as 137CsCl or 131CsCl, caesium chloride is used in nuclear medicine applications such as treatment of cancer and diagnosis of myocardial ...
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Sucrose
Sucrose, a disaccharide, is a sugar composed of glucose and fructose subunits. It is produced naturally in plants and is the main constituent of white sugar. It has the molecular formula . For human consumption, sucrose is extracted and refined from either sugarcane or sugar beet. Sugar mills – typically located in tropical regions near where sugarcane is grown – crush the cane and produce raw sugar which is shipped to other factories for refining into pure sucrose. Sugar beet factories are located in temperate climates where the beet is grown, and process the beets directly into refined sugar. The sugar-refining process involves washing the raw sugar crystals before dissolving them into a sugar syrup which is filtered and then passed over carbon to remove any residual colour. The sugar syrup is then concentrated by boiling under a vacuum and crystallized as the final purification process to produce crystals of pure sucrose that are clear, odorless, and sweet. Suga ...
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Percoll
Percoll is a tool for efficient density separation in Cell biology that was first formulated by Pertoft and colleagues. It is used for the isolation of cells, organelles, and/or viruses by density centrifugation. Percoll consists of colloidal silica particles of 15–30 nm diameter (23% w/w in water) which have been coated with polyvinylpyrrolidone (PVP). Percoll is well suited for density gradient experiments because it possesses a low viscosity compared to alternatives, a low osmolarity, and no toxicity towards cells and their constituents. Percoll is a registered trademark of GE Healthcare. Past use in artificial reproduction Percoll was previously used in assisted reproductive technology (ART) to select sperm from semen by density gradient centrifugation, for use in techniques such as ''in vitro'' fertilization or intrauterine insemination. However, in 1996, Pharmacia sent out a letter to laboratories stating that Percoll should be used for research purposes only, no ...
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CsCl Density Gradient Centrifugation
CSCL can refer to: * Caesium chloride (CsCl), a chemical compound. * Computer Supported Collaborative Learning, a research topic on supporting collaborative learning with the assistance of computer artifacts. * China Shipping Container Lines, a containerized marine shipping company, based in Shanghai Shanghai (; , , Standard Mandarin pronunciation: ) is one of the four direct-administered municipalities of the People's Republic of China (PRC). The city is located on the southern estuary of the Yangtze River, with the Huangpu River flowin ...
China. {{disambig ...
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GC Content
In molecular biology and genetics, GC-content (or guanine-cytosine content) is the percentage of nitrogenous bases in a DNA or RNA molecule that are either guanine (G) or cytosine (C). This measure indicates the proportion of G and C bases out of an implied four total bases, also including adenine and thymine in DNA and adenine and uracil in RNA. GC-content may be given for a certain fragment of DNA or RNA or for an entire genome. When it refers to a fragment, it may denote the GC-content of an individual gene or section of a gene (domain), a group of genes or gene clusters, a non-coding region, or a synthetic oligonucleotide such as a primer. Structure Qualitatively, guanine (G) and cytosine (C) undergo a specific hydrogen bonding with each other, whereas adenine (A) bonds specifically with thymine (T) in DNA and with uracil (U) in RNA. Quantitatively, each GC base pair is held together by three hydrogen bonds, while AT and AU base pairs are held together by two hydrogen bonds. ...
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Satellite DNA
Satellite DNA consists of very large arrays of tandemly repeating, non-coding DNA. Satellite DNA is the main component of functional centromeres, and form the main structural constituent of heterochromatin. The name "satellite DNA" refers to the phenomenon that repetitions of a short DNA sequence tend to produce a different frequency of the bases adenine, cytosine, guanine, and thymine, and thus have a different density from bulk DNA such that they form a second or "satellite" band(s) when genomic DNA is separated along a cesium chloride density gradient using buoyant density centrifugation. Sequences with a greater ratio of A+T display a lower density while those with a greater ratio of G+C display a higher density than the bulk of genomic DNA. Some repetitive sequences are ~50% G+C/A+T and thus have buoyant densities the same as bulk genomic DNA. These satellites are called "cryptic" satellites because they form a band hidden within the main band of genomic DNA. "Isopycnic" is ...
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Isopycnic
An isopycnic surface is a surface of constant density inside a fluid. Isopycnic surfaces contrast with isobaric or isothermal surfaces, which describe surfaces of constant pressure and constant temperature respectively. Isopycnic surfaces are sometimes referred to as "iso-density" surfaces, although this is strictly incorrect. Isopycnic typically describes surfaces, not processes. Unless there is a flux of mass into or out of a control volume, a process which occurs at a constant density also occurs at a constant volume and is called an isochoric process and not an isopycnic process. The term "isopycnic" is commonly encountered in the fluid dynamics of compressible fluids, such as in meteorology and geophysical fluid dynamics, astrophysics, or the fluid dynamics of explosions or high Mach number flows. It may also be applied to other situations where a continuous medium has smoothly varying density, such as in the case of an inhomogeneous colloidal suspension. In general isopycni ...
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Satellite DNA
Satellite DNA consists of very large arrays of tandemly repeating, non-coding DNA. Satellite DNA is the main component of functional centromeres, and form the main structural constituent of heterochromatin. The name "satellite DNA" refers to the phenomenon that repetitions of a short DNA sequence tend to produce a different frequency of the bases adenine, cytosine, guanine, and thymine, and thus have a different density from bulk DNA such that they form a second or "satellite" band(s) when genomic DNA is separated along a cesium chloride density gradient using buoyant density centrifugation. Sequences with a greater ratio of A+T display a lower density while those with a greater ratio of G+C display a higher density than the bulk of genomic DNA. Some repetitive sequences are ~50% G+C/A+T and thus have buoyant densities the same as bulk genomic DNA. These satellites are called "cryptic" satellites because they form a band hidden within the main band of genomic DNA. "Isopycnic" is ...
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Separation Processes
A separation process is a method that converts a mixture or a solution of chemical substances into two or more distinct product mixtures, a scientific process of separating two or more substance in order to obtain purity. At least one product mixture from the separation is enriched in one or more of the source mixture's constituents. In some cases, a separation may fully divide the mixture into pure constituents. Separations exploit differences in chemical properties or physical properties (such as size, shape, mass, density, or chemical affinity) between the constituents of a mixture. Processes are often classified according to the particular properties they exploit to achieve separation. If no single difference can be used to accomplish the desired separation, multiple operations can often be combined to achieve the desired end. With a few exceptions, elements or compounds exist in nature in an impure state. Often these raw materials must go through a separation before th ...
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