Bio-17-ATP (Biotin-17-adenosine-5’-triphosphate) can replace ATP for in vitro transcription reaction catalyzed by T3, T7 or SP6 RNA polymerases. The biotin-labeled RNA transcripts produced by these reactions are suitable for a wide range of applications such as nucleic acid hybridization, sequencing, and genome analysis.
The transcription reaction produces multiple RNA copies of the DNA template(s) during a short incubation period. RNA probes offer higher target specificity and greater sensitivity than the corresponding DNA-DNA hybrids. The single-stranded RNA probes offer slectivity unavailable with double-stranded DNA probes, because they are strand-specific. Furthermore, RNA probes hybridize much more efficiently to target molecules than DNA probes because there is no self-hybridization. The biotin-labeled hybridized probes can be detected by a reporter molecule linked to streptavidin, avidin, or anit-biotin antibody. Such a complex can be detected directly, e.g. by excitation of a fluorophore conjugated to streptavidin, or indirectly, e.g. using an enzyme conjugate that can produce an insoluble colored precipitate.
