Electrophoretic Color Marker
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Electrophoretic Color Marker
An electrophoretic color marker is used to monitor the progress of agarose gel electrophoresis and polyacrylamide gel electrophoresis (PAGE) since DNA, RNA, and most proteins are colourless. They are also referred to as tracking dyes, and are frequently present in loading dyes as well as molecular weight ladders. Examples Commonly used color markers include Bromophenol blue, Cresol Red, Orange G and Xylene cyanol. Generally speaking, Orange G migrates faster than bromophenol blue, which migrates faster than xylene cyanol, but the apparent "sizes" of these dyes (compared to DNA molecules) varies with the concentration of agarose and the buffer system used. For instance, in a 1% agarose gel made in TAE buffer (Tris-acetate-EDTA), xylene cyanol migrates at the speed of a 3000 base pair (bp) molecule of DNA and bromophenol blue migrates at 400 bp. However, in a 1% gel made in TBE buffer (Tris-borate A borate is any of several boron oxyanions, negative ions consisting of boron and oxy ...
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Agarose Gel Electrophoresis
Agarose gel electrophoresis is a method of gel electrophoresis used in biochemistry, molecular biology, genetics, and clinical chemistry to separate a mixed population of macromolecules such as DNA or proteins in a matrix of agarose, one of the two main components of agar. The proteins may be separated by charge and/or size (isoelectric focusing agarose electrophoresis is essentially size independent), and the DNA and RNA fragments by length. Biomolecules are separated by applying an electric field to move the charged molecules through an agarose matrix, and the biomolecules are separated by size in the agarose gel matrix. Agarose gel is easy to cast, has relatively fewer charged groups, and is particularly suitable for separating DNA of size range most often encountered in laboratories, which accounts for the popularity of its use. The separated DNA may be viewed with stain, most commonly under UV light, and the DNA fragments can be extracted from the gel with relative ease. ...
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Polyacrylamide Gel Electrophoresis
Polyacrylamide gel electrophoresis (PAGE) is a technique widely used in biochemistry, forensic chemistry, genetics, molecular biology and biotechnology to separate biological macromolecules, usually proteins or nucleic acids, according to their electrophoretic mobility. Electrophoretic mobility is a function of the length, conformation, and charge of the molecule. Polyacrylamide gel electrophoresis is a powerful tool used to analyze RNA samples. When polyacrylamide gel is denatured after electrophoresis, it provides information on the sample composition of the RNA species. Hydration of acrylonitrile results in formation of acrylamide molecules () by nitrile hydratase. Acrylamide monomer is in a powder state before addition of water. Acrylamide is toxic to the human nervous system, therefore all safety measures must be followed when working with it. Acrylamide is soluble in water and upon addition of free-radical initiators it polymerizes resulting in formation of polyacrylamide. ...
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Bromophenol Blue
Bromophenol blue (3′,3″,5′,5″-tetrabromophenolsulfonphthalein, BPB), albutest is used as a pH indicator, an electrophoretic color marker, and a dye. It can be prepared by slowly adding excess bromine to a hot solution of phenolsulfonphthalein in glacial acetic acid. Acid–base indicator As an acid–base indicator, its useful range lies between pH 3.0 and 4.6. It changes from yellow at pH 3.0 to blue at pH 4.6; this reaction is reversible. Bromophenol blue is structurally related to phenolphthalein (a popular indicator). Color marker Bromophenol is also used as a colour marker to monitor the process of agarose gel electrophoresis and polyacrylamide gel electrophoresis. Since bromophenol blue carries a slight negative charge at moderate pH, it will migrate in the same direction as DNA or protein in a gel; the rate at which it migrates varies according to gel density and buffer composition, but in a typical 1% agarose gel in a 1X TAE buffer or TBE buff ...
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Cresol Red
Cresol red (full name: ''o''-cresolsulfonephthalein) is a triarylmethane dye frequently used for monitoring the pH in aquaria. Molecular biology Cresol red can be used in many common molecular biology reactions in place of other loading dyes. Cresol Red does not inhibit ''Taq'' polymerase to the same degree as other common loading dyes. Color marker Cresol red can also be used as an electrophoretic color marker to monitor the process of agarose gel electrophoresis and polyacrylamide gel electrophoresis. In a 1% agarose gel, it runs approximately at the size of a 125 base pair (bp) DNA molecule (it depends on the concentration of buffer and other component). Bromophenol blue and xylene cyanol Xylene cyanol can be used as an electrophoretic color marker, or tracking dye, to monitor the process of agarose gel electrophoresis and polyacrylamide gel electrophoresis. Bromophenol blue and orange G Orange G also called C.I. 16230, Acid ... can also be used for this purpose. ...
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Orange G
Orange G also called C.I. 16230, Acid Orange 10, or orange gelb is a synthetic azo dye used in histology in many staining formulations. It usually comes as a disodium salt. It has the appearance of orange crystals or powder. Staining Orange G is used in the Papanicolaou stain to stain keratin. It is also a major component of the Alexander test for pollen staining. It is often combined with other yellow dyes and used to stain erythrocytes in the trichrome methods. Color marker Orange G can be used as an electrophoretic color marker to monitor the process of agarose gel electrophoresis, running approximately at the size of a 50 Base pair (bp) DNA molecule, and polyacrylamide gel electrophoresis. Bromophenol blue and xylene cyanol Xylene cyanol can be used as an electrophoretic color marker, or tracking dye, to monitor the process of agarose gel electrophoresis and polyacrylamide gel electrophoresis. Bromophenol blue and orange G Orange G also called C.I. 16230, Acid . ...
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Xylene Cyanol
Xylene cyanol can be used as an electrophoretic color marker, or tracking dye, to monitor the process of agarose gel electrophoresis and polyacrylamide gel electrophoresis. Bromophenol blue and orange G Orange G also called C.I. 16230, Acid Orange 10, or orange gelb is a synthetic azo dye used in histology in many staining formulations. It usually comes as a disodium salt. It has the appearance of orange crystals or powder. Staining Orange G ... can also be used for this purpose. Once mixed with the sample, the concentration of xylene cyanol is typically about 0.005% to 0.03%. Migration speed In 1% agarose gels, xylene cyanol migrates at about the same rate as a 4 to 5 kilobase pair DNA fragment,{{cite book , author = Lela Buckingham and Maribeth L. Flaws , title = Molecular Diagnostics: Fundamentals, Methods, & Clinical Applications , url = https://archive.org/details/moleculardiagnos00lela , url-access = limited , publisher = F.A. Davis Company , date = 2007 , pag ...
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Buffer Solution
A buffer solution (more precisely, pH buffer or hydrogen ion buffer) is an aqueous solution consisting of a mixture of a weak acid and its conjugate base, or vice versa. Its pH changes very little when a small amount of strong acid or base is added to it. Buffer solutions are used as a means of keeping pH at a nearly constant value in a wide variety of chemical applications. In nature, there are many living systems that use buffering for pH regulation. For example, the bicarbonate buffering system is used to regulate the pH of blood, and bicarbonate also acts as a buffer in the ocean. Principles of buffering Buffer solutions resist pH change because of a chemical equilibrium between the weak acid HA and its conjugate base A−: When some strong acid is added to an equilibrium mixture of the weak acid and its conjugate base, hydrogen ions (H+) are added, and the equilibrium is shifted to the left, in accordance with Le Chatelier's principle. Because of this, the hydrogen io ...
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Agarose
Agarose is a heteropolysaccharide, generally extracted from certain red seaweed. It is a linear polymer made up of the repeating unit of agarobiose, which is a disaccharide made up of D-galactose and 3,6-anhydro-L-galactopyranose. Agarose is one of the two principal components of agar, and is purified from agar by removing agar's other component, agaropectin. Agarose is frequently used in molecular biology for the separation of large molecules, especially DNA, by electrophoresis. Slabs of agarose gels (usually 0.7 - 2%) for electrophoresis are readily prepared by pouring the warm, liquid solution into a mold. A wide range of different agaroses of varying molecular weights and properties are commercially available for this purpose. Agarose may also be formed into beads and used in a number of chromatographic methods for protein purification. Structure Agarose is a linear polymer with a molecular weight of about 120,000, consisting of alternating D-galactose and 3,6-anhydro- ...
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Tris
Tris, or tris(hydroxymethyl)aminomethane, or known during medical use as tromethamine or THAM, is an organic compound with the formula (HOCH2)3CNH2, one of the twenty Good's buffers. It is extensively used in biochemistry and molecular biology as a component of buffer solutions such as in TAE and TBE buffers, especially for solutions of nucleic acids. It contains a primary amine and thus undergoes the reactions associated with typical amines, e.g. condensations with aldehydes. Tris also complexes with metal ions in solution. In medicine, tromethamine is occasionally used as a drug, given in intensive care for its properties as a buffer for the treatment of severe metabolic acidosis in specific circumstances. Some medications are formulated as the "tromethamine salt" including Hemabate (carboprost as trometamol salt), and "ketorolac trometamol". Buffering features The conjugate acid of tris has a p''K''a of 8.07 at 25 °C, which implies that the buffer has an effective pH r ...
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Acetate
An acetate is a salt (chemistry), salt formed by the combination of acetic acid with a base (e.g. Alkali metal, alkaline, Alkaline earth metal, earthy, Transition metal, metallic, nonmetallic or radical Radical (chemistry), base). "Acetate" also describes the conjugate acid, conjugate base or ion (specifically, the negatively charged ion called an anion) typically found in aqueous solution and written with the chemical formula . The neutral molecules formed by the combination of the acetate ion and a ''positive'' ion (called a cation) are also commonly called "acetates" (hence, ''acetate of lead'', ''acetate of aluminum'', etc.). The simplest of these is hydrogen acetate (called acetic acid) with corresponding salts, esters, and the polyatomic ion, polyatomic anion , or . Most of the approximately 5 billion kilograms of acetic acid produced annually in industry are used in the production of acetates, which usually take the form of polymers. In nature, acetate is the most common ...
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EDTA
Ethylenediaminetetraacetic acid (EDTA) is an aminopolycarboxylic acid with the formula H2N(CH2CO2H)2sub>2. This white, water-soluble solid is widely used to bind to iron (Fe2+/Fe3+) and calcium ions (Ca2+), forming water-soluble complexes even at neutral pH. It is thus used to dissolve Fe- and Ca-containing scale as well as to deliver iron ions under conditions where its oxides are insoluble. EDTA is available as several salts, notably disodium EDTA, sodium calcium edetate, and tetrasodium EDTA, but these all function similarly. Uses Textile industry In industry, EDTA is mainly used to sequester (bind or confine) metal ions in aqueous solution. In the textile industry, it prevents metal ion impurities from modifying colours of dyed products. In the pulp and paper industry, EDTA inhibits the ability of metal ions, especially Mn2+, from catalysing the disproportionation of hydrogen peroxide, which is used in chlorine-free bleaching. In a similar manner, EDTA is added ...
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Borate
A borate is any of several boron oxyanions, negative ions consisting of boron and oxygen, such as orthoborate , metaborate , or tetraborate ; or any salt with such anions, such as sodium metaborate, and disodium tetraborate . The name also refers to certain functional groups in molecules consisting of boron and oxygen, and esters with such groups, such as triethyl orthoborate . Natural occurrence Borate ions occur, alone or with other anions, in many borate and borosilicate minerals such as borax, boracite, ulexite (boronatrocalcite) and colemanite. Borates also occur in seawater, where they make an important contribution to the absorption of low frequency sound in seawater. Borates also occur in plants, including almost all fruits. Anions The main borate anions are: * tetrahydroxyborate , found in sodium tetrahydroxyborate . * orthoborate , found in trisodium orthoborate * perborate , as in sodium perborate * metaborate or , found in sodium metaborate * diborate , f ...
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