Enzyme Explorer

Nucleases



Purification of proteins and specific nucleic acids often requires the digestion of DNA, RNA or both. Viscosity problems resulting from high DNA concentrations and enzymatic cell dissociation methods are often enhanced utilizing DNAse.

Sigma offers a complete selection of high-purity nucleases to meet most digestion requirements.


      DNA DNA   Hybrid    
Non-Specific Nucleases Endo-nuclease Exo-nuclease Single
Strand
Double Strand RNA RNA Yields 3'-Phosphate Yields 5'-Phosphate
Benzonase X   X X X     X
DNAse I X   X X       X
DNAse II X   X X     X  
Exonuclease III   X   X       X
Micrococcal Nuclease X X X X     X  
Nuclease P1     X   X     X
Nuclease S1 X X X   X     X
Phosphodiesterase I   X           X
Phosphodiesterase II   X X X X     X
RNAse A X       X   X  
RNAse H X         X   X
RNAse T1 X       X   X  
Ribonuclease Optimized Blend X       X X X  
 
Restriction Endonucleases

Benzonase®

Endonuclease, recombinant from Serratia marcescens expressed in E.coli. (3.1.30.2)

Hydrolyzes double- and single-stranded DNA and RNA yielding 5'-phosphomononucleotide and 5'-phosphooligonucleotide end-products. Optimum temperature is 37 °C with operating range of 0-40 °C. It is active in the presence of ionic and nonionic detergents, reducing agents and urea.
Used for the removal of nucleic acids from protein samples and for the reduction of viscocity due to nucleic acids.


Product #  Product Name Add to Cart
E1014 Benzonase ≥250 units/μL, ≥90% (SDS-PAGE), recombinant, expressed in Escherichia coli, buffered aqueous glycerol solution
E8263 Benzonase, ultrapure ≥250 units/μL, ≥99% (SDS-PAGE), recombinant, expressed in Escherichia coli, buffered aqueous glycerol solution, ultrapure grade

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Deoxyribonuclease I

from bovine pancreas (3.1.21.1)

Catalyzes the endonucleolytic cleavage of double and single stranded DNA to yield 5'-phosphodinucleotide and 5'-phosphooligonucleotide end-products. The product of hydrolysis is a complex mixture of 5'-phosphate mononucleotides and oligonucleotides. In the presence of magnesium ions, DNase I attacks each strand of DNA independently and the cleavage sites are random. In the presence of manganese(II), DNase I cleaves both strands of DNA at approximately the same site. Most protocols use magnesium ion with DNase I but for specific purposes, manganese is used.

Molecular weight: 30,072 calculated from sequence.
Structure: a single polypeptide containing two disulfide bridges and a single carbohydrate moiety of monosaccharides.
Optimum pH for activity: 7-8
A protease-free DNase is stable at pH 5-7 up to 60 °C for at least five hours; at 62 °C, a 1 mg/mL solution lost activity at 6% per hour in either acetate buffer (pH 5) or Tris buffer (pH 7.2). Activity was destroyed at 68 °C. (The same research indicated that a solution at 0.1 mg/mL showed no activity loss after five hours at 62 °C.)


Product #  Product Name Add to Cart
D5025 Deoxyribonuclease I from bovine pancreas Type IV, lyophilized powder, Protein ~90 %, ≥2,000 Kunitz units/mg protein
D4527 Deoxyribonuclease I from bovine pancreas Type II, lyophilized powder, Protein ~90 %, ≥2,000 units/mg protein
D4513 Deoxyribonuclease I from bovine pancreas Type II-S, lyophilized powder, Protein ~90 %, ≥2,000 units/mg protein
D4263 Deoxyribonuclease I from bovine pancreas Standardized vial containing 2,000 Kunitz units of DNase I (D4527).
DN25 Deoxyribonuclease I from bovine pancreas lyophilized powder, Protein ≥85 %, 400-800 Kunitz units/mg protein
DNEP Deoxyribonuclease I from bovine pancreas lyophilized powder, Protein ~80 %, ≥1,500 Kunitz units/mg protein
D7691 Deoxyribonuclease I from bovine pancreas from bovine, recombinant, expressed in proprietary host
D7291 Deoxyribonuclease I RNase-free solution from bovine pancreas buffered aqueous glycerol solution

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Deoxyribonuclease II

from bovine spleen (3.1.21.1)

Catalyzes the endonucleolytic cleavage of double and single stranded DNA to yield nucleoside 3'-phosphates and 3'-phosphooligonucleotide end-products.

Molecular weight: 38,000 Da
The pH range for activity is 4.0 to 6.5, with only about 15% at pH 6.5. The optimum is 5.0. The optimum stability of the enzyme is at pH 5 - 5.5, with rapid inactivation at pH 8.5 at 30 °C.


Product #  Product Name Add to Cart
D8764 Deoxyribonuclease II from bovine spleen Type V, essentially salt-free, lyophilized powder, ≥1,000 units/mg protein
D4138 Deoxyribonuclease II from porcine spleen Type IV, lyophilized powder, 2,000-6,000 Kunitz units/mg protein (biuret)

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Exonuclease III

Catalyzes the exonucleolytic cleavage in the 3'- to 5'-direction to yield nucleoside 5'-phosphates. Has a preference for double-stranded DNA. May exhibit endonuclease activity at apurinic sites on DNA. (3.1.30.2)

Product #  Product Name Add to Cart
E1131 Exonuclease III from Escherichia coli BE25 /psGR3 buffered aqueous glycerol solution

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Micrococcal Nuclease

from Staphylococcus aureus (3.1.31.1)
Catalyzes the exo- and endonucleolytic cleavage of both double and single stranded DNA and RNA, with preference for AT and AU-rich regions, yielding nucleoside 3'-phosphates and 3'-phosphooligonucleotide end-products. It is most active against single stranded DNA.

Molecular Weight: 16,807
This enzyme has an absolute need for Ca2+ for activity.
The optimal pH is between 9 and 10, depending on the Ca2+ concentration. At higher pH values, less Ca2+ is required. The inhibitory effect of high Ca2+ concentrations is more pronounced at higher pH values. Mg2+ cannot replace Ca2+ in activating the enzyme.



Product #  Product Name Add to Cart
N3755 Nuclease micrococcal from Staphylococcus aureus 100-300 units/mg protein
N5386 Nuclease micrococcal from Staphylococcus aureus 100-300 units/mg protein

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Nuclease P1

from Penicillium citrinum (3.1.30.1)
Catalyzes the nonspecific endonucleolytic cleavage of single stranded DNA and RNA to yield nucleoside 5'-phosphates and 5'-phosphooligonucleotides. It does not appreciably degrade double-stranded nucleic acids, especially in the presence of more than 400 mM sodium chloride at pH 6.0.

Molecular Weight: 42-50 kDa.
A zinc dependent glycoprotein consisting of 270 amino acid residues.
The enzyme has an optimal temperature of approximately 70 °C, but for a long incubation, a temperature below 60 °C is more suitable. It is stable in the pH range of 5 - 8.



Product #  Product Name Add to Cart
N8630 Nuclease P1 from Penicillium citrinum lyophilized powder, ≥200 units/mg protein (E1%/280, 3'-5'-Phosphodiesterase)

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Nuclease S1

from Aspergillus oryzae (3.1.30.1)
Catalyzes the nonspecific endo- and exonucleolytic cleavage of single stranded DNA and RNA to yield nucleoside 5'-phosphates and 5'-phosphooligonucleotides. It is used to digest non-annealed polynucleotide tails and hairpin loops in RNA and DNA duplexes and can be used to convert super-helical DNA to the linear form.

Molecular weight: approximately 34 kDa and exists as a monomer.
The optimal pH range is 4.0 to 4.6 and it is activated by Zn2+ and/or Ca2+ ions. It is inhibited by chelators, such as EDTA and citrate, and by high concentrations of SDS.

Nuclease S1 may exhibit some nickase activity at high concentrations. This nickase activity can be inhibited by including a high concentration of NaCl (approximately 200 mM) in the reaction buffer.



Product #  Product Name Add to Cart
N5661 Nuclease S1 from Aspergillus oryzae

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Phosphodiesterase I

(3.1.4.1)
Catalyzes the exonucleolytic cleavage in the 3'- to 5'-direction to yield nucleoside 5'-phosphates. Low activity towards polynucleotides. A 5'-phosphate terminus on the substrate inhibits hydrolysis.



Product #  Product Name Add to Cart
P4631 Phosphodiesterase I from Bothrops atrox Type V, crude dried venom, ≥0.01 unit/mg solid
P3243 Phosphodiesterase I from Crotalus adamanteus venom 1 vial of ≥0.4 units, Purified
P3134 Phosphodiesterase I from Crotalus adamanteus venom Type VI, crude dried venom, ≥0.01 unit/mg solid
P4506 Phosphodiesterase I from Crotalus atrox (Western Diamondback Rattlesnake) Type IV, crude dried venom, ≥0.01 unit/mg solid

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Phosphodiesterase II

from bovine spleen (3.1.16.1 formerly 3.1.4.18)
Catalyzes the exonucleolytic cleavage of 5'-hydroxy-terminated oligonucleotides in the 5'- to 3'-direction to yield nucleoside 3'-phosphates. Preference is for a single-stranded substrate.



Product #  Product Name Add to Cart
P9041 Phosphodiesterase II from bovine spleen lyophilized powder, ≥5 units/mg protein

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Ribonuclease A

(3.1.27.5)
Catalyzes the endonucleolytic cleavage of RNA to yield nucleoside 3'-phosphates and 3'-phosphooligonucleotides ending in Cp or Up.

Molecular weight: 13,700
Activators of RNase A include potassium and sodium salts. The optimal temperature for activity is 60 °C, although the enzyme does exhibit activity from 15-70 °C. The pH optimum is 7.6, with an activity range of 6-10. The highest activity is exhibited with single stranded RNA. RNase A is a very stable enzyme and can withstand temperatures up to 100 °C. At 100 °C, RNase A is most stable between pH 2.0 and 4.5.


Product #  Product Name Add to Cart
R6513 Ribonuclease A from bovine pancreas for molecular biology, ≥70 Kunitz units/mg protein, lyophilized powder
R4642 Ribonuclease A from bovine pancreas for molecular biology, ≥70 Kunitz units/mg protein, buffered aqueous glycerol solution
R4875 Ribonuclease A from bovine pancreas Type I-A, powder, ≥60% as RNase A (SDS-PAGE), 50-100 Kunitz units/mg protein
R5503 Ribonuclease A from bovine pancreas Type I-AS, 50-100 Kunitz units/mg protein
R5000 Ribonuclease A from bovine pancreas Type II-A, ≥60% (SDS-PAGE), 50-100 Kunitz units/mg solid
R5125 Ribonuclease A from bovine pancreas Type III-A, ≥85% (SDS-PAGE), 85-140 Kunitz units/mg protein
R5250 Ribonuclease A from bovine pancreas Type X-A, ≥90% (SDS-PAGE), ~100 Kunitz units/mg protein
R5500 Ribonuclease A from bovine pancreas Type XII-A, ≥90% (SDS-PAGE), ~100 Kunitz units/mg protein

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Ribonuclease H

from E. coli ( 3.1.4.34)
Specifically hydrolyzes the phosphodiester bonds of RNA in RNA:DNA duplexes to generate products with 3'-hydroxyl and 5'-phosphate ends. It degrades only the RNA component of the DNA-RNA hybrid (RNA that is hydrogen bonded to a complementary DNA strand).

Molecular Weight: 17.6 kDa
The pH optimum for ribonuclease H is 7.5 to 9.1. The enzyme is activated by Mg2+ (2 - 4 mM). RNase H is a sulfhydryl containing enzyme, and is activated by the presence of dithiothreitol and inhibited by N-ethylmalemide.


Product #  Product Name Add to Cart
R6501 Ribonuclease H from Escherichia coli recombinant, expressed in Escherichia coli, buffered aqueous glycerol solution, 1,000-4,000 units/mL

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Ribonuclease T1

(3.1.27.1)
Catalyzes the two-stage endonucleolytic cleavage of RNA to yield nucleoside 3'-phosphates and 3'-phosphooligonucleotides ending mainly in Gp. In the reaction, cleavage occurs between the 3'-phosphate group of a guanidine ribonucleotide and 5'-hydroxyl of the adjacent nucleotide. The initial product is a 2':3' cyclic phosphate nucleoside that is hydrolyzed to the corresponding 3'-nucleoside phosphate.

Molecular Weight: 11,068
In solution, it is fairly resistant to heat (100 °C for 10 minutes at pH 6) and acid, but unstable in alkaline solution (>pH 9). It should be noted that the reaction catalyzed by the enzyme can not be stopped by heating the reaction mixture to 100 °C.


Product #  Product Name Add to Cart
R1003 Ribonuclease T1 from Aspergillus oryzae ammonium sulfate suspension, 300,000-600,000 units/mg protein

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