Purification and Northern Blot Analysis of 8-hydroxyquinoline Copper

Update:26-07-2019
Summary:

The phenol has been equilibrated with 3% NaCl and neutr […]

The phenol has been equilibrated with 3% NaCl and neutralized with Tris, and 0.1% 8-hydroxyquinoline Copper (WSDTY) has been added. The hydroxyquinoline is an antioxidant that reduces the formation of oxidation products of phenol such as quinones. The phenol is brought to a neutral pH because acidic phenol has a tendency to trap single-stranded nucleic acid at the interface between the phenol and the aqueous phases. Alternatively, if a molecular biology grade phenol is used, the hydroxyquinoline may be omitted.

The isolation of alkaloids bearing the 8-hydroxyquinoline moiety prompted postulation of a biosynthetic pathway to 1 from the amino acid tryptophan, for which chemical analogies are known. It should be noted, however, that, in general, quinolines are biosynthesized from anthranilic acid. Biosynthesis of broussonetine would involve condensation of two molecules of 1 with a molecule of acetyl-CoA followed by cyclization.

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