Investigation of 9-Anthryldiazomethane as a fluorescent label for nucleotides, by Antoinette L. Vergallito .

Abstract

Currently, biotin is the most used nonradioactive label for nuc1eotides. This paper examines the use of9-anthryldiazomethane (ADAM) as an alternate method for labeling nucleotides, extending its use as a fluorescent labeling reagent to include phosphorylated as well as carboxyl containing biomolecules. Adenosine 5'-monphosphate (AMP) was the primary nucleotide investigated. Studies were also conducted on the synthesis and purification ofADAM. A wide variety oftechniques were used in this project. Thin layer chromatography was used to monitor reactions. Attempts to separate the mixture after synthesis ofADAM utilized gravity, flash, and high pressure liquid chromatography. Mass spectrometry and 31p NMR were used to test for the presence of an ADAM-AMP derivative.