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S1 Nuclease

#EN0321 Supplied with: 5X Reaction Buffer	10,000 u (100 u/µl) 2x1 ml
Related Documents (in pdf, ~40-60 KB): Certificate of Analysis: #EN0321 MSDS (English) MSDS (English-USA) MSDS (German)

Description
The S1 Nuclease degrades single-stranded nucleic acids, releasing 5'-phosphoryl mono- or oligonucleotides. It is five times more active on DNA than on RNA (1). The S1 Nuclease exhibits 3'-phosphomonoesterase activity.
The enzyme is a glycoprotein with carbohydrate content of 18%.

Source
Aspergillus oryzae.

Molecular Weight
29 kDa monomer.

Applications

• 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), see Protocol for Generation of Unidirectional Deletions in DNA Fragments.

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).

Concentration
100 u/µl

Definition of Activity Unit
One unit of the enzyme produces 1 µg 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, 800 µg/ml heat denatured calf thymus DNA.

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

5X Reaction Buffer
200 mM sodium-acetate (pH 4.5 at 25°C), 1.5 M NaCl and 10 mM ZnSO₄.

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.

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

Related Products

Exonuclease III, E.coli Water, nuclease-free

References

1. The Enzymes, 3rd. Ed. (Boyer, P.D., ed.), Endonucleases specific for single-stranded polynucleotides, Lehman, R.I.; review), 4,193-201,1981.
2. Roberts T.M. et al., A general method for maximizing the expression of a cloned gene, Proc. Natl. Acad. Sci. USA, 76, 760-764, 1979.
3. Berk, A.J., Sharp, P.A., Spliced early mRNAs of simian virus 40, Proc. Natl. Acad. Sci. USA, 75, 1274-1278, 1978.
4. Weidle, U., Weissmann, C., The 5'-flanking region of a human IFN-alpha gene mediates viral induction of transcription, Nature, 303, 442-446, 1983.
5. Henikoff, S., Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing, Gene, 28, 351-359, 1984.
6. Vogt, V.M., Purification and further properties of single-strand-specific nuclease from Aspergillus oryzae, Eur. J. Biochem., 33, 192-200, 1973.

Updated vasario 05, 2008 14:35