DNA FISH probe

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DNA FISH probe can replace TTP in reactions where it serves as a substrate for E. coli DNA polymerase 1 (holoenzyme and Klenow fragment), terminal deoxynucloetide transferase, T4 and Taq DNA polymerases, and reverse transcriptases (from AMV and M-MuLV). 波长：Ex (nm) 454,Em(nm) 511
DNA FISH probe is incorporated into DNA by a variety of labeling reactions including nick translation, random primed DNA synthesis and 3’ end labeling reactions. Allylamine-labeled DNA can be efficiently conjugated to the active ester form of signal generating moieties such as fluorescent dyes, biotin and other haptens to produce labeled probes for hybridization/detection assays. The efficient incorporation of Allylamine-dUTP, in contrast to many dye-labeled nucleotides, provides for greater incorporation of signaling moieties and stronger signals.

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DNA FISH probe（allylamine-dUTP ）优点：
Using indirect labeling with allyamine-dUTPs provide great advantages for FISH probe synthesis that may not be present in direct labeling. One key benefit is that virtually any reporter molecule containing a reactive amine group such as NHS can be linked to the probe. While this of course means molecules such as biotin and other haptens can potentially bind, in the case of FISH it means that virtually any fluorescent molecule can be attached. This provides a high degree of flexibility that can be tailored to one’s own preferences, the same of which cannot be said for direct labeling. While it is true that direct labeling requires fewer steps and can sometimes be cheaper, there are significant hindrances in nucleotide incorporation. Depending on bulkiness, pre-conjugated fluorescent nucleotides can induce steric effects as well as block active and binding sites in transcriptional machinery. Allylamine-dUTPs, on the other hand, do not have these same drawbacks. In various studies, allylamine-dUTP transcription has been shown to have almost identical transcriptional efficiency compared to unmodified NTPs. When conjugating allylamine-dUTP probes with amine-reactive fluorescent molecules in excess, one can expect high levels of fluorophore incorporation and consistency regardless of the chosen fluorescent dye.

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