Technical Data
D3935
DNA Polymerase I
500u
2500u
2500u
|
Technical Data |
|
D3935 |
DNA Polymerase I |
500u 2500u |
| Cloning | Storage: -20°CShipping: Blue Ice |
|
DNA Polymerase I catalyzes the polymerization of nucleotides into duplex DNA in the 5’=>3’ direction. The enzyme exhibits 3’-5’ proofreading exonuclease activity and 5’=>3’ degrading activity. Purified from an E.coli strain carrying a DNA polymerase I overproducing plasmid. Application: • Second strand cDNA synthesis • Nick translation Supplied with: D3935-05: DNA Polymerase I Reaction Buffer, 10X Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer. Molecular Weight: 109kD |
Source: E. coli, strain carrying a DNA polymerase I overproducing plasmid. Form: Supplied as a liquid in PBS, pH 7.5, 1mM DTT, 50% glycerol. Concentration: 10u/ul D3935-05, Reaction Buffer (10X): Supplied as a liquid in 500mM Tris-HCl, pH 7.5 (25°C), 100mM MgCl2, 10mM DTT Unit Definition: One unit of enzyme will catalyze the exchange of 10nmole of total DNA to a DE-81 adsorbable form in 30 minutes at 37°C using poly[dA-dT)] as a template primer. Quality Control: Endodeoyuribonuclease Assay: No detectable conversion of covalently closed circular DNA to nicked DNA was observed after incubation of 20u of DNA Ploymerase with 1ug of pBR322 DNA in 50ul of reaction buffer for 4 hours at 37°C. Activity Assay: 67mM potassium phosphate (pH 7.4), 6.7mM MgCl2, 1mM 2-mercaptoethanol, 0.033mM dATP, 0.033mM dTTP, 0.4MBq/ml [3H]-dTTP and 62.5ug/ml poly(dA-dT)•poly(dA-dT). Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. |
|
1. W. S. Kelly and K. H. Stump, J. Biol. Chem., 254: 3206 (1968). 2. T. Maniatis, et al., Proc. Natl. Acad. Sci. USA, 72: 1184 (1975). 3. N. E. Murray and W. S. Kelly, Molec. Gen Genet., 175: 77 (1979). 4. Gubler, U., Hoffmann, B.J., Gene 25: 263-2369 (1983). 5. Current Protocols in Molecular Biology, vol1 (Ausubel, F.M., et al., ed.), John Wiley and Sons, Inc., Brooklyn, NY, 3.5.3-3.5.6, (1994-2001).
|
||