Compound for sequencing by synthesis

摘要

The present invention provides deoxynucleoside tri- or tetraphosphate comprising a 3′ nitrate and a detectable label covalently bound to the oxygen atom of an oxymethyl or oxyallyl or oxypropargyl substitution of a nucleobase. Such compounds provide new possibilities for future Sequencing by Synthesis technologies.

主权项

1. A method for analyzing a DNA sequence comprising the steps of
a) providing a nucleic acid template with a primer hybridized to said template forming a primer/template hybridization complex, b) adding DNA polymerase, and at least one labeled deoxynucleoside tri- or tetraphosphate comprising a 3′-nitrate and a detectable label covalently bound to the oxygen atom of an oxymethyl or oxyallyl or oxypropargyl substitution of a nucleobase, and a pyrophosphatase, c) subjecting said reaction mixture to conditions which enable a DNA polymerase catalyzed primer extension reaction, d) determining, whether said compound has been incorporated into said primer/template hybridization complex, e) simultaneously removing the 3′-nitrate group and optionally said label from said complex by means of adding a sulfide or a mercaptane, and f) repeating steps b) to e) at least once.