| Description |
T7 DNA Polymerase catalyzes DNA synthesis in the 5'->3' direction. It is a highly processive DNA polymerase allowing continuous synthesis of long stretches of DNA. The enzyme also exhibits a very strong 3'->5' exonuclease activity towards single- and double-stranded DNA.
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| Applications |
- Purification of covalently closed circular DNA by removal of residual genomic DNA
- Primer extension reactions on long templates
- DNA 3'-end labeling
- Strand extensions in site-directed mutagenesis
- Fill-in blunting of 5'-overhang DNA
- Second strand synthesis of cDNA
- In situ detection of DNA fragmentation associated with apoptosis
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| Source |
T7 DNA Polymerase is composed of two subunits: an 80 kDa polypeptide (the product of gene 5 of bacteriophage T7) and a 12 kDa thioredoxin from the trxA gene of E. coli. Each protein is cloned and overexpressed separately in E. coli. |
| Unit Definition |
One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid-precipitable material in 30 minutes at 37°C. |
| Components |
- T7 DNA Polymerase: 10,000 U/ml in 50 mM KPO4, 0.1 mM EDTA, 1.0 mM DTT, 50% glycerol, pH 7.0 @ 25°C
- 10X Reaction Buffer: 200 mM Tris-HCl, 100 mM MgCl2 10 mM DTT, pH 7.5 @ 25°C
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| Quality Control |
The absence of endodeoxyribonucleases is confirmed by the appropriate quality test. |
| Storage Condition |
-20 °C |