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Chloroform: Chloroform is stabilized with small quantities of amylene or ethanol, because exposure of pure chloroform to oxygen and ultraviolet light produces phosgene gas. Some chloroform solutions come as pre-made a 96% chloroform, 4% isoamyl alcohol mixture that can be mixed with an equal volume of phenol to obtain the 25:24:1 solution.
Aqueous samples, lysed cells, or homogenised tissue are mixed with equal volumes of a phenol:chloroform mixture. This mixture is then centrifuged. Because the phenol:chloroform mixture is immiscible with water, the centrifuge will cause two distinct phases to form: an upper aqueous phase, and a lower organic phase.
Isoamyl alcohol is a colorless liquid with the formula C 5 H 12 O, specifically (H 3 C–) 2 CH–CH 2 –CH 2 –OH. It is one of several isomers of amyl alcohol (pentanol). It is also known as isopentyl alcohol, isopentanol, or (in the IUPAC recommended nomenclature) 3-methyl-butan-1-ol. An obsolete name for it was isobutyl carbinol. [5]
This page contains tables of azeotrope data for various binary and ternary mixtures of solvents. The data include the composition of a mixture by weight (in binary azeotropes, when only one fraction is given, it is the fraction of the second component), the boiling point (b.p.) of a component, the boiling point of a mixture, and the specific gravity of the mixture.
Phenol is often used in combination with chloroform. [4] Adding an equal volume of chloroform and phenol ensures a distinct separation between the aqueous and organic phases. Chloroform and phenol are miscible and create a denser solution than phenol alone, aiding the separation of the organic and aqueous layers. This addition of chloroform is ...
or isoamyl alcohol or isopentyl alcohol primary 3-Methylbutan-1-ol: 131.2 2,2-dimethyl-1-propanol or neopentyl alcohol primary 2,2-Dimethylpropan-1-ol: 113.1 2-pentanol or sec-amyl alcohol or methyl (n) propyl carbinol secondary Pentan-2-ol: 118.8 3-methyl-2-butanol or sec-isoamyl alcohol or methyl isopropyl carbinol secondary 3-Methylbutan-2 ...
The specific method used to extract the DNA, such as phenol-chloroform extraction, alcohol precipitation, or silica-based purification. [4] For the chemical method, many different kits are used for extraction, and selecting the correct one will save time on kit optimization and extraction procedures.
Phenol is also a versatile precursor to a large collection of drugs, most notably aspirin but also many herbicides and pharmaceutical drugs. Phenol is a component in liquid–liquid phenol–chloroform extraction technique used in molecular biology for obtaining nucleic acids from tissues or cell culture samples.