One-lane chemical sequencing of 3'-fluorescent-labeled DNA.
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.
Ferraboli
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Dipartimento di Scienze Biomediche e Biotechnologie, Università di Brescia, Italy.
Negri
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R
Di Mauro
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Barlati
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D001322:Autoanalysis; D001483:Base Sequence; D003199:Computer Systems; D004247:DNA / Q000737:chemistry*; D003720:Densitometry; D005453:Fluorescence; D005559:Formamides; D007834:Lasers; D008969:Molecular Sequence Data; D010880:Piperidines; D017354:Point Mutation
D005559:Formamides; D010880:Piperidines; C031066:formamide; C032727:piperidine; D004247:DNA