Nukleasen

Enzyme

Substrate

Activity

Products

Applications

Heat Inactivation

Exonuclease III,
 E. coli
(162025)

dsDNA

3´→5´ exonuclease that digests one strand of duplex DNA from the 3´ end of a nick or a blunt or 3´-recessed end; not active on thionucleotides. Exo III also has 3´-DNA phosphatase and apurinic DNA endonuclease activities.

dNMPs and ssDNA on the opposite strand. Partial digestion produces dsDNA having 5´ extensions of ssDNA.

Used with S1 Nuclease or Mung Bean Nuclease to make nested deletions. Preparation of ssDNA templates for sequencing. Site-directed mutagenesis. Preparation of labeled strand-specific probes.

20 min, 70°C

RNase-Free DNase I (bovine pancreas, 170500, 171000)

dsDNA and ssDNA

Activated by divalent cations. In presence of Mg²⁺, it cleaves each DNA strand randomly and independently, preferentially adjacent to pyrimidines. In presence of Mn²⁺, it cleaves both strands simultaneously, generating fragments with blunt ends of 1-2-base overhangs

Oligos and dNMPs with 5´ phosphate and 3´-OH

Removing DNA from RNA preps. Random nicking of dsDNA. DNase footprinting

20 min, 75°C

Baseline-ZERO™ Dnase (170005)

dsDNA and ssDNA

Digests dsDNA or ssDNA down to mononucleotides

Mononucleotides

Removing DNA from RNA preps

20 min, 75°C

Plasmid-Safe™ ATP-Dependent Dnase (161010, 161100)

linear ssDNA and dsDNA

Selectively digests linear DNA. No activity on nicked or closed-circular DNA.

dNMPs

Remove chromosomal DNA fragments from plasmid, fosmid, and BAC preparations.

NT

Exonuclease I, E. coli (162200)

ssDNA

3´→5´ exonuclease,  it requires the presence of magnesium and a free 3´-hydroxyl terminus; wide range of buffer conditions 

dNMPs

Removal of ssDNA and oligonucleotides.

15 min, 80°C

Exonuclease VII
(162225)

ssDNA

Exonuclease that digests in both 5´→3´ and 3´→5´ directions.

dNMPs

Removal of primers and single-stranded oligos.

10 min, 95°C

Rec J Exonuclease (162337)

ssDNA

5´→3´ exonuclease that digests ssDNA with a 5´ phosphate or a 5´OH.

dNMPs

Removal of primers and ssDNA from dsDNA.

20 min, 65°C

Terminator™ 5´-Phosphate-Dependent Exonuclease
(162370)

ssDNA or ssRNA

5´→3´ exonuclease that digests ssDNA or ssRNA with 5´-monophosphorylated ends, but not with 5´-hydroxyl, 5´-triphosphorylated, or 5´-capped ends

dNMPs or NMPs

Removal of 5´-monophosphorylated DNA or primers or oligos. Enrichment of ssDNA or ssRNA molecules lacking 5´-monophosphate groups

10 min, 65°C

OmniCleave™ Endonuclease
(129350)

ssDNA, dsDNA, or RNA

Endonuclease that efficiently digests DNA and RNA

di-, tri-, and tetra-nucleotides

Removal of DNA and RNA from protein preps. Removal of host DNA from phage preps

No

Hybridase™ Thermostable RNase H
(108211)

RNA in RNA:DNA hybrid

Cleaves RNA in RNA:DNA hybrid without affecting unhybridized RNA or DNA.

Oligoribonucleotides with 5´ phosphate and 3´ OH

High-stringency hybrid selection.

No

RNase A
(160630)

ssRNA

Cleaves ssRNA 3´ of pyrimidine residues.

Oligoribonucleotides with 3´-cytidine or 3´-uridine residues

Removal of RNA from DNA preps. RNase protection assays. RNA mapping and structure studies.

NT

RNase I, E. coli
(173010)

ssRNA

Cleaves ssRNA between all dinucleotide pairs.

NMPs with 5´-OH and 2´,3´-cyclic monophosphate

Removal of RNA from DNA preps. RNase protection assays. Mismatch detection of single basepairs in RNA:RNA or RNA:DNA hybrids.

15 min, 70°C

RNase R
(172010)

linear RNA-not circular or lariat

Magnesium-dependent 3´→5´ exoribonuclease 

Alternative splicing and gene expression studies ,Intron cDNA production, Intronic screening of cDNA libraries