| Criteria | RNase A | RNase T1 | Remarks |
| Full Name | Ribonuclease A | Ribonuclease T1 | Both are ribonucleases but differ in sequence specificity and cleavage pattern |
| Substrate Specificity | Cleaves single-stranded RNA at the 3′ end of pyrimidine bases (C, U) | Cleaves single-stranded RNA at the 3′ end of guanosine residues | RNase A targets pyrimidines, RNase T1 targets guanosines |
| Cleavage Site | After cytosine or uracil residues | After guanine residues | Site-specific cleavage differs based on the nucleotide base preference |
| Source | Bovine pancreas | Aspergillus oryzae (fungus) | RNase A is animal-derived; RNase T1 is fungal-derived |
| Enzyme Type | Endoribonuclease | Endoribonuclease | Both cleave RNA internally but at different sequence contexts |
| Optimal Conditions | pH ~7.0, requires no metal ions | pH ~7.5, requires no metal ions | Both operate under mild, similar lab conditions |
| Structural Features | Small, stable, resistant to denaturation | Single-chain enzyme with tight sequence specificity | RNase T1 is more selective, RNase A more broadly acting |
| Applications in Research | RNA degradation, RNA footprinting, sample clean-up | RNA sequencing, RNA structure analysis, RNA digestion mapping | RNase T1 is especially useful for site-specific cleavage in sequencing or structural probing |
| Inhibition Sensitivity | Sensitive to RNase inhibitors (e.g., RNasin) | Not affected by RNase A inhibitors | Inhibitor selection matters when working with combinations of RNases |
| Use in RNA Mapping | Useful for generating RNA ladders, non-specific cleavage | Ideal for base-specific cleavage pattern for RNA sequencing | RNase T1 provides finer resolution in RNA structure-function studies |
