DNA Polymerase I Klenow Fragment CAS 9012-90-2

CAS Number

9012-90-2

Grade

Molecular Biology Grade

Molecular Weight

EU Commodity Code

38220090

Shipping Temp

Blue Ice

Storage Temp

-20°C

E. coli lysogenic CM 5199

DNA Polymerase I Large Fragment exhibits the 5’ ---> 3’ polymerase and 3’ ---> 5’ exonuclease activity of DNA Polymerase. Lacks 5’ ---> 3’ exonuclease activity.

Applications

• Random primer labeling • Sanger sequencing • cDNA Second Strand Synthesis • 3’ end-labeling, filling • Blunting of DNA ends: fill-in 5'-overhangs or/and removal of 3'-overhangs, see protocol • Oligonucleotide-directed site-specific mutagenesis.

Source

E. coli cells with a cloned fragment of the polA gene

Concentration

~10u/ul

Storage Buffer

Supplied as a liquid in 25mM Tris-HCl, pH 7.5, 0.1mM EDTA, 1mM DTT, 50% glycerol.

Unit Definition

1u of DNA Polymerase I Klenow Fragment catalyzes the incorporation of 10nmoles of deoxynucleotides into polynucleotide fraction in 30 minutes at 37°C.

D3940A DNA Polymerase I Klenow Fragment, Reaction Buffer, 10X

Supplied as a liquid in 500mM Tris-HCl, pH 8.0, 50mM magnesium chloride, 10mM DTT

Inhibition and Inactivation

-Inhibitors: metal chelators, nucleotide analogs PPi, Proprietray information (at high concentrations). -Inactivated by heating at 75°C for 10 min or by addition of EDTA.

Endodeoxyribonuclease Assay

No detectable degradation was observed after incubation of supercoiled plasmid DNA with DNA Polymerase I Klenow Fragment.

Storage and Stability

Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 6 months after receipt 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.

Source

E. coli cells with a cloned fragment of the polA gene

Concentration

~10u/ul

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.

References

1. C.C. Richardson, et al., J. Biol. Chem., 239:222 (1964) 2. F. Sanger, et al., Proc. Natl. Acad. Sci., 74: 5463 (1977).3. J. C. Telford, et al., Proc. Natl. Acad. Sci., 76: 2590 (1979). 4. Current Protocols in Molecular Biology, vol. 1 (Ausubel, F.M., et al., eds.), John Wiley & Sons, Inc. Brooklyn, NY, 3.5.7-3.5.10, 1994-2001. 5. Rougeon, F., et al., Nucleic Acids Res., 2:2365-2378, 1975. 6. Feinberg, et al., Anal. Biochem., 137:266-267, 1984

USBio References

No references available

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D3940 DNA Polymerase I Klenow Fragment CAS: 9012-90-2

CAS Number

Grade

Molecular Weight

EU Commodity Code

Shipping Temp

Storage Temp

E. coli lysogenic CM 5199

References

USBio References

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