5-bromo-2′-deoxy-uridine labeled nucleotide triphosphates and nucleic acid probes and methods of making and using the same
View Patent ↗5-bromo-2′-deoxy-uridine (BrdU) labeled nucleotide triphosphates and nucleic acid probes are described herein. The BrdU labeled nucleotide triphosphates include a linker between the nucleotide triphosphate and the BrdU moiety. The linker can be cleavable or non-cleavable. The nucleotide triphosphates can be a ribonucleotide triphosphates, 2′-deoxyribonucleotide triphosphates or 2′,3′-dideoxyribonucleotide triphosphates. The nucleic acid probes can be used for in situ hybridization.
1. A compound of the formula:
or a salt thereof, wherein “Nu” is a nucleotide triphosphate, a 2′-deoxynucleotide triphosphate or a 2′,3′-dideoxynucleotide triphosphate and wherein L is a linker.
2. The compound of claim 1 , wherein L has a length greater than a C 11 n-alkane carbon chain.
3. The compound of claim 1 , wherein “Nu” is uridine triphosphate, 2′-deoxyuridine triphosphate or 2′,3′-dideoxyuridine triphosphate.
4. The compound of claim 1 , wherein the compound is of the formula:
or a salt thereof, wherein each X is independently H or OH and “L” is a linker.
5. The compound of claim 1 , wherein “L” is a linker having a chemical structure selected from the group consisting of:
wherein n is an integer of from 6 to 12.
6. The compound of claim 1 , wherein the compound is of the formula:
or a salt thereof, wherein each X is independently H or OH.
7. A polynucleotide represented by the formula:
wherein “Y” is a nucleoside, a nucleotide or a polynucleotide, “Z” is H, a nucleotide or a polynucleotide, “X” is H or OH, “B” is a nucleobase and “L” is a linker.
8. The polynucleotide of claim 7 , wherein the polynucleotide is represented by the formula:
wherein “Y” is a nucleoside, nucleotide or polynucleotide, “Z” is H, a nucleotide or a polynucleotide, “X” is H or OH and “L” is a linker.
9. The polynucleotide of claim 7 , wherein “L” is a linker having a chemical structure selected from the group consisting of:
wherein n is an integer of from 6 to 12.
10. A polynucleotide represented by the formula:
wherein “Y” is a nucleoside, nucleotide or polynucleotide, “Z” is H, a nucleotide or a polynucleotide and “X” is H or OH.
11. A method for detecting the presence of a nucleic acid analyte in a tissue sample comprising:
incubating the tissue sample with a composition comprising the polynucleotide of claim 7 , wherein the polynucleotide binds to the nucleic acid analyte;
washing the sample;
contacting the sample with an anti-BrdU antibody conjugated to a detectable moiety or to an enzyme; and
detecting the detectable moiety or enzyme.
12. A composition comprising:
adenosine triphosphate (ATP);
uridine triphosphate (UTP);
guanosine triphosphate (GTP);
cytidine triphosphate (CTP); and
a compound as set forth in claim 1 , wherein “Nu” is a nucleotide triphosphate.
13. A kit comprising:
a composition as set forth in claim 12 ; and
an RNA polymerase.
14. A composition comprising:
dATP, dCTP, dGTP, dTTP and a compound as set forth in claim 1 , wherein “Nu” is a 2′-deoxynucleotide triphosphate or a 2′,3′-dideoxynucleotide triphosphate.
15. A kit comprising:
a composition as set forth in claim 14 , wherein “Nu” is a 2′-deoxynucleotide triphosphate; and
a DNA polymerase.
16. A kit comprising:
a composition as set forth in claim 14 , wherein “Nu” is a 2′,3′-dideoxynucleotide triphosphate; and
terminal transferase.
17. A sodium salt of the compound of claim 1 .
18. A sodium salt of the compound of claim 6 .
19. The method of claim 11 , wherein the sample comprises a fixed sample.