5-bromo-2′-deoxy-uridine labeled nucleotide triphosphates and nucleic acid probes and methods of making and using the same
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 method comprising:
reacting a compound (1) which is bound to a support:
wherein “R 1 ” is a group linking the nucleoside moiety to the support and “HPG” is a protecting group, with the compound (2):
wherein R 2 is a linking group and R 3 is H or SO 3 − , to form a compound (3):
reacting the compound (3) with the compound (4):
wherein each X is independently H or OH, to form a compound (5);
and removing the compound (5) from the support to form a compound (6);
2. The method of claim 1 , wherein R 2 is selected from the group consisting of:
wherein n is an integer of from 6 to 12.
3. The method of claim 1 , wherein R 1 comprises an amino acid.
4. The method of claim 3 , wherein R 1 comprises:
wherein R 4 is an amino acid.
5. The method of claim 1 , wherein “Y” is a dimethoxytrityl (DMT) group.