Thermo Scientific™ Nucleasi S1

Descrizione

S1 Nuclease degrades single-stranded nucleic acids, releasing 5'-phosphoryl mono- or oligonucleotides. It is five times more active on DNA than on RNA . S1 Nuclease also cleaves dsDNA at the single-stranded region caused by a nick, gap, mismatch or loop. S1 Nuclease exhibits 3'-phosphomonoesterase activity.

The enzyme is a glycoprotein with carbohydrate content of 18%.

• Source: Aspergillus oryzae
• Molecular Weight: 29 kDa monomer

• The absence of contaminating double-stranded DNA specific nuclease activity confirmed by appropriate quality test
• Functionally tested for the generation of unidirectional deletions in DNA fragments (in conjunction with Exonuclease III)

• Aspergillus oryzae

• 29 kDa monomer

• One unit of the enzyme produces 1μg of acid soluble deoxyribonucleotides in 1 min. at 37°C
• Enzyme activity is assayed in the following mixture: 30 mM sodium acetate (pH 4.5), 50 mM NaCl, 0.1 mM ZnCl₂, 5% (v/v) glycerol, 800μg/ml heat denatured calf thymus DNA

• The enzyme is supplied in:20mM Tris-HCl (pH 7.5), 50mM NaCl, 0.1mM ZnCl₂ and 50% (v/v) glycerol

• 200mM sodium acetate (pH 4.5 at 25°C), 1.5M NaCl and 10 mM ZnSO₄

• Inhibitors: metal chelators, PP_i, P_i, 5'-ribonucleotides and deoxyribonucleotides
• Inactivated by heating at 70°C for 10 min. in the presence of EDTA

Recommended for:

Removal of single-stranded overhangs of DNA fragments (2); S1 transcript mapping (3, 4); Cleavage of hairpin loops; Creation of unidirectional deletions in DNA fragments in conjunction with Exonuclease III (5)

Note:

S1 Nuclease can introduce breaks into double-stranded DNA, RNA and DNA/RNA hybrids at high enzyme and low salt concentrations (6).