Fluorophores and haptens, the latter meaning Biotin, Desthiobiotin, Digoxigenin and Dinitrophenol, are the most commonly used labels for the generation of non-radioactive RNA/cRNA probes. They are enzymatically introduced into RNA/cRNA via modified nucleotides that are incorporated as substitutes for their natural counterparts in either a one-step procedure (introduction of a fluorescent or hapten-modified nucleotide) or a two-step procedure (introduction of a reactive group carrying nucleotide and subsequent fluorescent or hapten labeling).
Nucleotide Selector for RNA Labeling
Select a suitable nucleotide substrate for enzymatic RNA Labeling.
| Label | Labeling Kits | Single Nucleotides |
|---|---|---|
| Fluorophore | Fluorescent in vitro Transcription Kits | Fluorescent Nucleotides |
| Hapten | Biotin&Digoxigenin in vitro Transcription Kits | Biotinylated Nucleotides Digoxigenin-/DNP-modified Nucleotides |
| Amine | Amine-modified Nucleotides | |
| CLICK | Copper-free CLICk T7 in vitro Transcription Labeling Kits | CLICK-functionalized Nucleotides |
| Nucleotide dependent |
| Template | Method | Labeled Probe | Labeling Site | Enzyme |
|---|---|---|---|---|
| DNA | in vitro Transcription | cRNA | random | T7- / SP6- / T3-RNA Polymerase |
| RNA | 3’ End Labeling | RNA | 3’ OH | Terminal deoxynucleotidyl Transferase (TdT) |
| RNA | 3’ OH | Yeast Poly A Polymerase | ||
| RNA | 3’ OH | T4 RNA Ligase | ||
| 5’ End Labeling | RNA | 5’ OH | T4 Polynucleotide Kinase (T4 PNK) |
