Exonuclease I
Detailní popis
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).
Highlights
- Active in PCR buffers
Applications
- 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)
Note
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 |
|
Hazardous | No |
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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