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One-lane chemical sequencing of 3'-fluorescent-labeled DNA
S Ferraboli1, R Negri, E Di Mauro
1Dipartimento di Scienze Biomediche e Biotechnologie, Università di Brescia, Italy.
Analytical Biochemistry
|November 1, 1993
Abstract:
A method for DNA sequencing that combines limited chemical degradation of 3'-fluorescent-labeled DNA with densitometric analysis using an automatic sequencer is presented. The DNA sequence is determined in a single electrophoretic lane by monitoring the intensities of bands representing products of cleavage at the four bases obtained by solvolysis in hot aqueous piperidine (10%) followed by treatment with hot formamide. An application of the method for the detection of point mutations is reported.