Nucleotide analogues
AssigneeQiagen Sciences, LLC
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.
Claims23
A labeled deoxynucleoside triphosphate according to the following structure:
- wherein D is selected from the group consisting of an azide, disulfide alkyl, disulfide substituted alkyl groups;
- wherein said label is a dye and B is a nucleobase.
A labeled deoxynucleoside triphosphate according to the following structure:
- wherein D is selected from the group consisting of an azide, disulfide alkyl, disulfide substituted alkyl groups;
- wherein said label is a dye and B is a nucleobase.
A labeled deoxynucleoside triphosphate according to the following structure:
- wherein D is selected from the group consisting of an azide, disulfide alkyl, disulfide substituted alkyl groups;
- wherein said label is a dye and B is a nucleobase.
A kit comprising a DNA polymerase and at least one deoxynucleoside triphosphate according to the following structure:
- wherein D is selected from the group consisting of an azide, disulfide alkyl, disulfide substituted alkyl groups, disulfide allyl, and disulfide substituted allyl groups;
- B is a nucleobase;
- A is an attachment group selected from the group consisting of exocyclic amine, propargyl amine, and propargyl hydroxyl;
- C is a cleavable site core selected from the group consisting of:
- and,
- wherein R1 and R2 are independently selected alkyl groups;
- L1 and L2, are connecting groups;
- and Label is a label selected from the group consisting of fluorophore dyes, energy transfer dyes, mass-tags, biotin, and haptenes.
A reaction mixture comprising a nucleic acid template with a primer hybridized to said template, a DNA polymerase and at least one deoxynucleoside triphosphate according to the following structure:
- wherein D is selected from the group consisting of an azide, disulfide alkyl, disulfide substituted alkyl groups, disulfide allyl, and disulfide substituted allyl groups;
- B is a nucleobase;
- A is an attachment group selected from the group consisting of exocyclic amine, propargyl amine, and propargyl hydroxyl;
- C is a cleavable site core selected from the group consisting of:
- wherein R1 and R2 are independently selected alkyl groups;
- L1 and L2, are connecting groups;
- and Label is a label selected from the group consisting of fluorophore dyes, energy transfer dyes, mass-tags, biotin, and haptenes.
Dependencies are inferred from the claim text.