Thermo Scientific Exonuclease I (ExoI) degrades single-stranded DNA in a 3'=>5' direction. It releases deoxyribonucleoside 5'-monophosphates in a stepwise manner and leaves 5'-terminal dinucleotides intact.
It does not cleave DNA strands with terminal 3'-OH groups blocked by phosphoryl or acetyl groups (see Reference 1).
- Active in PCR buffers
- Primer removal from PCR mixtures:
- prior to PCR product sequencing (see Reference 2)
- for one-tube "megaprimer" PCR mutagenesis (see Reference 3)
- Removal of single-stranded DNA containing a 3'-hydroxyl terminus from nucleic acid mixtures
- Assay for the presence of single-stranded DNA with a 3'-hydroxyl terminus (see Reference 4)
The enzyme is not suitable for removing 3'-overhangs of dsDNA.
|10X Reaction Buffer||670 mM glycine-KOH (pH 9.5 at 25°C), 67 mM MgCl2, 10 mM DTT.|
|Definition of Activity Unit||
|Inactivation||Inactivated by heating in boiling water bath for 10 min, preferably in the presence of EDTA.|
|Inhibition||Inhibitors: no specific inhibitors have been described.|
|Molecular Weight||54.5 kDa monomer.|
|Quality Control||The absence of other endo-, exodeoxyribonucleases, and ribonucleases confirmed by appropriate quality tests.|
|Source||E. coli cells with a cloned E. coli sbcB gene.|
|Storage Buffer||The enzyme is supplied in:
20 mM Tris-HCl (pH 7.5), 0.1 mM EDTA, 1 mM DTT and 50% (v/v) glycerol.
|Storage Condition||-20 C|
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