A novel biotinylated adenylate analogue derived from pyrazolo[3,4-d]pyrimidine for labeling DNA probes.
A novel dATP analogue, 3-[5-[(N-biotinyl-6- amiocaproyl)amino]pentyl]-1-(2-deoxy-beta-D-erythro-pentofuranosyl )-1H-pyrazolo[3,4-d]pyrimidin-4-amine 5'-triphosphate (9, bio-13-dAPPTP), which is modified at the 3-position with a flexible linker arm bearing a terminal biotin moiety, has been synthesized. This nucleotide is readily incorporated into DNA probes by nick translation. These probes hybridize to complementary targets as well as probes labeled with bio-dUTP, as judged by slot blot. When incorporated into oligonucleotides, they do not cause the loss of hybridization efficiency that an N-6-substituted adenine nucleotide does when incorporated into the same sites in the oligonucleotide.
Petrie
C R
CR
MicroProbe Corporation, Bothell, Washington 98021.
Adams
A D
AD
Stamm
M
M
Van Ness
J
J
Watanabe
S M
SM
Meyer
R B
RB
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000378:metabolism; D001483:Base Sequence; D001710:Biotin / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000378:metabolism; D004256:DNA Polymerase I / Q000378:metabolism; D015342:DNA Probes / Q000737:chemistry / Q000378:metabolism; D004279:DNA, Viral; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D009711:Nucleotides / Q000378:metabolism; D027383:Papillomaviridae / Q000235:genetics; D011720:Pyrazoles / Q000138:chemical synthesis* / Q000378:metabolism; D011743:Pyrimidines / Q000138:chemical synthesis* / Q000378:metabolism; D013696:Temperature
D015342:DNA Probes; D004279:DNA, Viral; D009711:Nucleotides; D011720:Pyrazoles; D011743:Pyrimidines; C074013:3-(5-((N-biotinyl-6-aminocaproyl)amino)pentyl)-1-(2-deoxyerythropentofuranosyl)-1H-pyrazolo(3,4-d)pyrimidin-4-amine-5'-triphosphate; D001710:Biotin; D000255:Adenosine Triphosphate; D004256:DNA Polymerase I