A: RNase T1 is a guanosine-specific endoribonuclease that cleaves single-stranded RNA at phosphodiester bonds immediately following guanosine (G) residues. It generates RNA fragments with 3′-phosphate and 5′-hydroxyl termini and does not degrade DNA.
A: RNase T1 preferentially cleaves single-stranded RNA. Guanosine residues located in double-stranded or highly structured regions of RNA are often protected from cleavage, making the enzyme useful for probing RNA secondary structure.
A: RNase T1 does not require divalent cations such as Mg²⁺ or Ca²⁺ for catalytic activity.
A: RNase T1 reactions are typically performed at neutral to slightly basic pH (approximately pH 7.0–8.0) using buffers such as Tris-HCl supplemented with EDTA to protect RNA from metal-dependent degradation.
A: EDTA is included to chelate divalent metal ions that could activate contaminating metal-dependent nucleases or promote RNA degradation. It helps preserve RNA integrity without inhibiting RNase T1 activity.
A: No. ScriptGuard™ RNase Inhibitor inhibits RNase A-family enzymes that cleave RNA at pyrimidines. RNase T1 belongs to a different enzyme family and is not affected.
A: RNase T1 provides base-specific cleavage, cutting only after guanosine residues. In contrast, non-specific RNases (such as RNase I or RNase A) cleave RNA at many or all nucleotide positions, resulting in complete RNA degradation.
A: RNase T1 degrades RNA without affecting DNA; however, because it cleaves only after guanosine residues, it may leave RNA fragments intact. For complete RNA removal, non-specific RNases are generally preferred.
A: RNase T1 activity units are defined by the manufacturer based on specific assay conditions. Users should follow product-specific unit definitions when calculating enzyme requirements.
