Exonuclease I, E. coli
Applications
Exonuclease I digests ssDNA in a 3´→5´ direction,1-3 but does not digest dsDNA. Although it requires the presence of magnesium and a free 3´-hydroxyl terminus for activity, it is active under a wide variety of buffer conditions and can be added directly into most reaction mixes. Exonuclease I can be heat-inactivated by incubation at 80°C for 15 minutes. Unit Definition: One unit of Exonuclease I catalyzes the release of 10 nmol of acid-soluble nucleotides from heat-denatured calf thymus DNA in 30 minutes at 37°C under standard assay conditions. Storage Buffer: 50% glycerol containing 50 mM Tris-HCl (pH 7.5), 0.1 M NaCl, 0.1 mM EDTA, 1 mM DTT, and 0.1% Triton® X-100. Quality Control: Exonuclease I is tested in degradation of ssDNA and is free of detectable RNase, endonuclease, and doublestranded exonuclease activities. References
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Figure 1. Specificity of Exonuclease I for ssDNA. 200 ng of EcoR I-linearized pUC19 dsDNA and 1 µg of a 100-mer ssDNA oligo were mixed in 1X TA Buffer and incubated at 37°C for 20 min in the absence or presence of 10 units of Exonuclease I. As seen on this 1% agarose gel, Exonuclease I completely digested the linear ssDNA oligo, but left the linearized plasmid dsDNA intact. Lane 1, size markers; Lane 2, minus Exo I; Lane 3, plus Exo I.
Ordering
Catalog No. | Concentration | Size |
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Exonuclease I, E. coli
X40501K | 20 U/µl | 1,000 Units |
X40505K | 20 U/µl | 5,000 Units |
X40520K | 20 U/µl | 20,000 Units |
Note: Catalogue number X40501K has been discontinued, effective from August 2014. This product is available only until stocks last.