Efficient incorporation of positively charged 2′, 3′-dideoxynucleoside-5′-triphosphates by DNA polymerases and their application in ‘direct-load’ DNA sequencing
AUTOR(ES)
Finn, Patrick J.
FONTE
Oxford University Press
RESUMO
A series of charge-modified, dye-labeled 2′, 3′-dideoxynucleoside-5′-triphosphates have been synthesized and evaluated as reagents for dye-terminator DNA sequencing. Unlike the commonly used dye-labeled terminators, these terminators possess a net positive charge and migrate in the opposite direction to dye-labeled Sanger fragments during electrophoresis. Post-sequencing reaction purification is not required to remove unreacted nucleotide or associated breakdown products prior to electrophoresis. Thus, DNA sequencing reaction mixtures can be loaded directly onto a separating medium such as a sequencing gel. The charge-modified nucleotides have also been shown to be more efficiently incorporated by a number of DNA polymerases than regular dye-labeled dideoxynucleotide terminators or indeed normal dideoxynucleoside-5′-triphosphates.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=169923Documentos Relacionados
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