Nucleotide compositions comprising photocleavable markers and methods of preparation thereof
First Claim
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1. A method of preparing a marker-photocleavable linker-nucleotide conjugate comprising:
- a) providing i) a photocleavable linker comprising a protective group, ii) an amino-nucleotide analog, and iii) an activated marker molecule selected from the group consisting of BODIPY-FL-SF and Cy5-NHS;
b) operably linking said photocleavable linker to said amino-nucleotide analog to produce a photocleavable linker-nucleotide conjugate;
c) removing said protective group from said photocleavable linker-nucleotide conjugate under conditions such that an activated photocleavable linker-nucleotide conjugate is created, wherein said activated photocleavable linker-nucleotide conjugate has an exposed reactive site on the linker portion of the conjugate; and
d) contacting said activated marker molecule with said activated photocleavable linker-nucleotide conjugate under conditions such that a marker-photocleavable linker-nucleotide conjugate is produced.
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Abstract
Labelled nucleotides and polynucleotides useful in the sequencing of nucleic acids are described. Methods of preparing photocleavable marker nucleotides and photocleavable marker-polynucleotide conjugates are described. Such photocleavable markere nucleotides can be incorporated into nucleic acid so as to create photocleavable marker-polynucleotide conjugates.
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11 Claims
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1. A method of preparing a marker-photocleavable linker-nucleotide conjugate comprising:
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a) providing i) a photocleavable linker comprising a protective group, ii) an amino-nucleotide analog, and iii) an activated marker molecule selected from the group consisting of BODIPY-FL-SF and Cy5-NHS; b) operably linking said photocleavable linker to said amino-nucleotide analog to produce a photocleavable linker-nucleotide conjugate; c) removing said protective group from said photocleavable linker-nucleotide conjugate under conditions such that an activated photocleavable linker-nucleotide conjugate is created, wherein said activated photocleavable linker-nucleotide conjugate has an exposed reactive site on the linker portion of the conjugate; and d) contacting said activated marker molecule with said activated photocleavable linker-nucleotide conjugate under conditions such that a marker-photocleavable linker-nucleotide conjugate is produced. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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11. A marker-photocleavable linker-nucleotide conjugate of the structure:
Specification