Acridinium ester labeling and purification of nucleotide probes

Details Acridinium ester labeling and purification of nucleotide probes Acridinium ester labeling and purification of nucleotide probes

C - Chemistry, Metallurgy

07

H

195/1.12

C07H 21/00 (2006.01)

Patent

CA 1314009

Aridinium Ester Labelling and Purification of Nucleotide Probes Abstract Methods for the construction, labelling and subsequent purification of nucleic acid probes containing primary amines with acridinium esters 4-(2- succinimidyloxycarbonyl-ethyl)phenyl10-methylacridinium- 9-carboxylate fluorosulfonate). The method for attaching acridinium esters to probes uses high (0.1 to 50mM) acridinium ester concentrations achieved using organic solvent in concentrations of 20% to 80% by volume, and may be carried out either in solution, or with one or the other of the acridinium ester or the probe suspended in solution. Purification (the separation of labelled probe from unlabelled probe and free label) involves (1) first removing most of the free acridinium ester label from probe using rapid separation techniques, then (2) removing substantially all remaining free label from the probe and separating labelled probe from unlabelled probe, involves specific applications of ion exchange, reverse phase or hydroxyapatite HPLC.

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