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Phenol: The phenol used for biochemistry comes as a water-saturated solution with Tris buffer, as a Tris-buffered 50% phenol, 50% chloroform solution, or as a Tris-buffered 50% phenol, 48% chloroform, 2% isoamyl alcohol solution (sometimes called "25:24:1"). Phenol is naturally somewhat water-soluble, and gives a fuzzy interface, which is ...
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. The aqueous phase rises to the top because it is less dense than the organic phase containing the phenol:chloroform.
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.
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]
Chloroform: 1.48 61.2 3.88 –63.5 ... 5.24 5.7 –7.00 Phenol: 181.75 3.60 43.0 ... [24] Dimethylacetamide: 166.1 [25] Tetrahydrofuran: 66.0
A commonly used method is guanidinium thiocyanate-phenol-chloroform extraction. It is not strictly necessary to use phenol or chloroform if extracting RNA for Northern blotting or DNA for Southern blot analysis because the gel electrophoresis followed by transfer to a membrane will separate the RNA/DNA from the proteins. Additionally, since ...
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