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US 11,548,909 B2Granted
Methods of using nucleotide analogues
AssigneeIsoPlexis Corporation
InventorsMong Sano Marma, Jerzy Olejnik, Ilia Korboukh
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Abstract
The present invention provides methods, compositions, mixtures and kits utilizing deoxynucleoside triphosphates comprising a 3′-O position capped by a group comprising methylenedisulfide as a cleavable protecting group and a detectable label reversibly connected to the nucleobase of said deoxynucleoside. Such compounds provide new possibilities for future sequencing technologies, including but not limited to Sequencing by Synthesis.
Claims2
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 a first deoxynucleoside triphosphate comprising a nucleobase and a sugar selected from a mixture of at least 4 differently labeled, 3′-O methylenedisulfide capped deoxynucleoside triphosphate compounds having the structures:
- c) subjecting said reaction mixture to conditions which enable a DNA polymerase catalyzed primer extension reaction so as to create a modified primer/template hybridization complex, and
- d) detecting said first detectable label of said labeled 3′-O methylenedisulfide capped deoxynucleoside triphosphate in said modified primer/template hybridization complex,
- e) removing said cleavable protecting group, and
- f) repeating steps b) to e) at least once.
3 more elements
Dependencies are inferred from the claim text.