Dideoxy DNA sequencing with end-labeled oligonucleotide primers |
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Authors: | R A McGraw |
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Affiliation: | Department of Biology, University of North Carolina, Chapel Hill, North Carolina 27514 USA |
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Abstract: | End-labeled oligonucleotide primers may be used effectively as the source of radiolabel in DNA sequencing by the dideoxy method. The approach is demonstrated with various end-labeled oligonucleotides, including a commercially prepared universal primer and a mixed-sequence probe. Single-stranded (M13) or denatured double-stranded template may be used. The end-labeled primers and their extension products may be stored for weeks. The method is useful for identification of clones isolated by oligonucleotide hybridization and it provides a convenient, economical alternative to the use of alpha-labeled deoxynucleoside triphosphate. |
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Keywords: | DNA sequencing dideoxy end-labeling oligonucleotide primer |
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