Get The DNase Activity Of RNase T And Its Application To DNA Cloning - Ncbi Nlm Nih

The function of nucleases (DNases and RNases) includes the enzymatic breakdown of DNA and RNA and is necessary for numerous research applications. For example, the purification of proteins and specific nucleic acids often requires the digestion of DNA, RNA or both.

Deoxyribonuclease (DNase) enzymes perform a variety of important cellular roles by degrading DNA via hydrolysis of its phosphodiester backbone. Deoxyribonuclease I (DNase I) enzymes cleave single or double-stranded DNA and require divalent metal ions to hydrolyze DNA yielding 3΄-hydroxyl and 5΄-phosphorylated products.

RNase A does not degrade DNA but can bind to DNA [25].

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.

RNases catalyze cleavage on RNA, which contributes to the degradation of RNA. They break the bonds between nucleotides, breaking RNA into smaller components and allowing access to other enzymes.

The function of nucleases (DNases and RNases) includes the enzymatic breakdown of DNA and RNA and is necessary for numerous research applications. For example, the purification of proteins and specific nucleic acids often requires the digestion of DNA, RNA or both.

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.

Ribonuclease A is a digestive enzyme secreted by the pancreas that specifically "digests" or hydrolyzes RNA (but not DNA) polymers by endonuclease cleavage of the phosphodiester bonds forming the covalent links between adjacent ribonucleotide residues in these molecules.

DNase activity is represented by the size of a dispensed circular well in an agarose gel layer, in which DNA stained by ethidium bromide is uniformly distributed. After the incubation, a circular dark zone is formed as the enzyme diffuses from the well radially into the gel and cleaves DNA.

We show that purified RNase T degrades ssDNA very efficiently, but under certain conditions, it is also able to completely degrade double-stranded (ds)DNA. Moreover, the action of RNase T on dsDNA with single-strand 3′-extensions can be controlled such that predominantly blunt-ended DNA is generated.

The function of nucleases (DNases and RNases) includes the enzymatic breakdown of DNA and RNA and is necessary for numerous research applications. For example, the purification of proteins and specific nucleic acids often requires the digestion of DNA, RNA or both.

Deoxyribonuclease (DNase) enzymes perform a variety of important cellular roles by degrading DNA via hydrolysis of its phosphodiester backbone. Deoxyribonuclease I (DNase I) enzymes cleave single or double-stranded DNA and require divalent metal ions to hydrolyze DNA yielding 3΄-hydroxyl and 5΄-phosphorylated products.

RNase A does not degrade DNA but can bind to DNA [25].

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.

RNases catalyze cleavage on RNA, which contributes to the degradation of RNA. They break the bonds between nucleotides, breaking RNA into smaller components and allowing access to other enzymes.

The function of nucleases (DNases and RNases) includes the enzymatic breakdown of DNA and RNA and is necessary for numerous research applications. For example, the purification of proteins and specific nucleic acids often requires the digestion of DNA, RNA or both.

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.

Ribonuclease A is a digestive enzyme secreted by the pancreas that specifically "digests" or hydrolyzes RNA (but not DNA) polymers by endonuclease cleavage of the phosphodiester bonds forming the covalent links between adjacent ribonucleotide residues in these molecules.

DNase activity is represented by the size of a dispensed circular well in an agarose gel layer, in which DNA stained by ethidium bromide is uniformly distributed. After the incubation, a circular dark zone is formed as the enzyme diffuses from the well radially into the gel and cleaves DNA.

We show that purified RNase T degrades ssDNA very efficiently, but under certain conditions, it is also able to completely degrade double-stranded (ds)DNA. Moreover, the action of RNase T on dsDNA with single-strand 3′-extensions can be controlled such that predominantly blunt-ended DNA is generated.