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.
|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)|