- Does not cleave DNA strands with terminal 3 ft.-OH groups blocked by phosphoryl or acetyl groups
- Active in PCR buffers
Quality Control:
- The absence of endo-, double-stranded exodeoxyribonucleases and ribonucleases confirmed by appropriate quality tests
Source:
- E.coli cells with a cloned E.coli sbcB gene
Molecular Weight:
Definition of Activity Unit:
- One unit of the enzyme catalyzes the release of 10nmol of acid soluble nucleotides in 30 min. at 37°C
- Enzyme activity is assayed in the following mixture: 67mM glycine-KOH (pH9.5), 6.7mM MgCl2, 1 mM DTT and 0.17mg/mL single-stranded [3H]-DNA
Storage Buffer:
- The enzyme is supplied in: 20mM Tris-HCl (pH7.5), 0.1mM EDTA, 1mM DTT and 50% (v/v) glycerol
10X Reaction Buffer:
- 670mM glycine-KOH (pH 9.5 at 25°C), 67mM MgCl2, 10mM DTT
Inhibition and Inactivation:
- Inhibitors: 20% (w/v) PEG 8000 (5)
- Inactivated by heating at 80°C for 15 min.
Recommended for: - Primer removal from PCR mixtures: prior to PCR product sequencing (2), for one-tube “megaprimer” PCR mutagenesis (3)
- Removal of single-stranded DNA containing a 3'-hydroxyl terminus from nucleic acid mixtures
- Assay for the presence of
The enzyme is not suitable for removing 3'-overhangs of dsDNA.
The purchase of this product allows the purchaser to use it for preparing amplified DNA fragments under a license from GE Healthcare of U.S. Patent Nos. 5,741,676 and 5,756,285 and other foreign patents.
5 ft. direction, releasing deoxyribonucleoside 5 ft.-monophosphates in a stepwise manner and leaving 5 ft.-terminal dinucleotides intact.
