Thermo Scientific™ S1 Nuclease (100 U/μL)

Description

• 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 Quality Control: 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) Source: Aspergillus oryzae Molecular Weight: 29 kDa monomer Definition of Activity Unit: 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 ZnCl 2 , 5% (v/v) glycerol, 800μg/ml heat denatured calf thymus DNA Storage Buffer: The enzyme is supplied in:20mM Tris-HCl (pH 7.5), 50mM NaCl, 0.1mM ZnCl 2 and 50% (v/v) glycerol 5X Reaction Buffer: 200mM sodium acetate (pH 4.5 at 25°C), 1.5M NaCl and 10 mM ZnSO 4 Inhibition and Inactivation: 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).