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RNA/cRNA Labeling

 

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 DNA/RNA Labeling
Select a suitable nucleotide substrate for enzymatic DNA/RNA Labeling.

 

Table 1: Standard enzymatic procedures for the preparation of non-radioactive RNA/cRNA probes.

TemplateMethodLabeled ProbeLabeling SiteEnzyme
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)