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  • RNases Offered by NEB

    RNA ReagentsReturn to RNases

    Exonuclease T (Exo T) (NEB #M0265) also known as RNase T, is a single-stranded RNA (1,2) or DNA (3,4) specific nuclease that requires a free 3´ terminus and removes nucleotides in the 3´→ 5´ direction. Exonuclease T can be used to generate blunt ends from RNA (5) or DNA molecules that have 3´ extensions (2).

    RNase H (Ribonuclease H) (NEB #M0297) is an endoribonuclease that specifically hydrolyzes the phosphodiester bonds of RNA which is hybridized to DNA. This enzyme does not digest single or double-stranded DNA. It can be used to remove poly(A) tails of mRNA hybridized to poly(dT), or to remove mRNA during second strand cDNA synthesis.

    Ribonuclease HII (RNase HII) (NEB #M0288) is an endoribonuclease that preferentially nicks 5´ to a ribonucleotide within the context of a DNA duplex. The enzyme leaves 5´ phosphate and 3´ hydroxyl ends (5). RNase HII will also nick at multiple sites along the RNA portion of an Okazaki fragment.

    ShortCut® RNase III (NEB #M0245) used with its manganese-containing reaction buffer, converts long double-stranded RNA into a heterogeneous mix of short (18–25 bp) interfering RNAs (siRNA) suitable for RNA interference in mammalian cells (6–8). 1.5 units (1 µl) of ShortCut RNase III is sufficient to convert 1 µg of dsRNA into siRNA suitable for RNA interference in mammalian cells.

    XRN-1 (NEB #M0338) is a highly processive 5´→3´ exoribonuclease, requiring 5´ monophosphate and can be used in removal of RNA containing 5´ monophosphate from a RNA mixture.

    Evaluation of RNase contamination is necessary for reagents to be used in experiments with RNA. The RNase Contamination Assay Kit detects general RNase activities, including non-enzyme based RNA degradation due to heavy metal contamination in samples and high pH.

    References

    1. Deutscher, M.P., Marlor, C.W. and Zaniewski, R. (1984) Proc. Natl. Acad. Sci. USA, 81, 4290-4293. PMID: 6379642
    2. Deutscher, M.P. and Marlor, C.W. (1985) J. Biol. Chem, 260, 7067-7071. PMID: 3888994
    3. Viswanathan, M., Dower, K. D. and Lovett, S. T. (1998) J. Biol. Chem, 273, 35126-35131. PMID: 9857048
    4. Zuo, Y. and Deutscher, M. P. (1999) Nucleic Acid Res, 27, 4077-4082. PMID: 10497273
    5. Rydberg, B. and Game, J. (2002) Proc. Natl. Acad. Sci., 99, 19954-16659. PMID: 12475934
    6. Morlighem, J.E. et al. (2007) Biotechniques, 42, 599-606. PMID: 17515198
    7. Yang, D. et al. (2002) Proc. Natl. Acad. Sci. USA, 99, 9942-9947. PMID: 12096193
    8. Calegari, F. et al. (2002) Proc. Natl. Acad. Sci. USA, 99, 14236-14240. PMID: 12391321