Modifying nucleotide analogues
AssigneeAmersham International plc
InventorsMichael Alan Reeve, Philip Steven Robinson
Life of this patent
Estimating the term…
Abstract
Fluorescently labelled dideoxynucleoside triphosphates are widely used as chain-terminators in Sanger dideoxy sequencing operations. But these unincorporated dye-terminators migrate in an electrophoresis gel and obscure the desired sequence ladder. This invention provides a method and a kit for modifying the unincorporated dye-terminators, e.g. by removal of a 5'-triphosphate group by chemical or enzymatic means e.g. by use of a phosphatase enzyme.
Claims8
A method of treating the product of a nucleic acid sequencing, mapping or labelling reaction containing one or more fluorescently-labeled nucleotides or analogues and DNA chains incorporating residues of the one or more fluorescently-labelled nucleotides or analogues to reduce the interference of unincorporated fluorescently-labelled nucleotides or analogues, which method comprises using an enzyme to remove at least one 5'-phosphate group of the one or more fluorescently-labelled nucleotides or analogues without correspondingly modifying the residues of the one or more fluorescently-labelled nucleotides or analogues incorporated in the DNA chains and then subjecting the treated product to gel electrophoresis.
Dependencies are inferred from the claim text.