Page id: 103

RNASE SUBSTRATE
 

Product description

RNase-substrate

 

The oligonucleotide 6-Fam-dArUdAdA-BHQ1 and the analogue 6-Fam-dArUdAdA-BMN-Q530 are used as a substrate for a convenient proof of RNases. In the inactivated state the quencher causes full extinction of the fluorescence of fluorescein. After digestion by the RNase, however, quencher and fluorophore are separated from each other and thus, the fluorescence of the 6-Fam reporter can be detected at 520 nm.

Schaubild der steigenden Fluoreszenz nach Spaltung des Substrats Fluoreszenzspektrum des gespaltenen Substrats
Increase of fluorescence induced by digestion of 6-Fam-dArUdAdA-BHQ1 (200 pM RNase A, 100 nM RNase substrats, 100 mM MES buffer, pH = 6.0, 100 mM NaCl), detection at 515 nm, excitation at 490 nm   
 
Fluorescene spectrum of the cleaved 6-Fam-dArUdAdA-BHQ1

Fluoreszenzfreisetzung nach RNase Substratspaltung
 
 


Literature:
1. Hypersensitive substrate for ribonucleases. Kelemen BR, Klink TA, Behlke MA, Eubanks SR, Leland PA, Raines RT; Nuc. Acids Chem. (1999), 27, 3696-3701.

2. A Ribonuclease Variant with Low Catalytic Activity but High Cytotoxicity. Bretscher LE, Abel RL, Raines RT.  J. Biol. Chem. (2000), 275.