| Product Name |
Klenow Fragment |
| Catalog Number |
EG-1055 |
| Description |
Klenow Fragment, a truncated form of Escherichia coli DNA Polymerase I expressed recombinantly in Escherichia coli, catalyzes 5’ to 3’ DNA synthesis and 3’ to 5’ proofreading exonuclease activity but lacks 5’ to 3’ exonuclease activity, suitable for blunt-end DNA synthesis and labeling applications. |
| Applications |
- Conversion of 3’ overhangs to blunt ends
- Random-primed DNA labeling
- Fill-in of 5’ overhangs of dsDNA to form blunt ends
- DNA sequencing by the Sanger method
- Site-directed mutagenesis with synthetic oligonucleotides
- Second-strand cDNA synthesis
|
| Source |
Recombinant Escherichia coli expressing the Klenow Fragment of the polA gene |
| Unit Definition |
One unit is defined as the amount of enzyme required to incorporate 10 nmol of dNTPs into acid-insoluble material in 30 minutes at 37°C under standard DNA polymerase assay conditions. |
| Components |
- Klenow Fragment: 5,000 units/mL in 25 mM Tris-HCl (pH 7.4 at 25°C), 1 mM DTT, 0.1 mM EDTA, 50% (v/v) glycerol
- 10X Reaction Buffer: 500 mM NaCl, 100 mM Tris-HCl (pH 7.9 at 25°C), 100 mM MgCl2, 10 mM DTT
|
| Quality Control |
- Absence of endodeoxyribonucleases confirmed by appropriate quality tests
- Functionally validated for fill-in of 5’ overhangs of dsDNA and random-primed DNA labeling
|
| Storage |
-20°C |
| Shipping |
Dry ice |