DNA Polymerase I (E. coli) 

DNA Polymerase I is a multifunctional enzyme that combines a DNA polymerase activity, a 5´→ 3´exonuclease activity and a 3’→5′ proofreading exonuclease activity. The 5´→3´ exonuclease activity enables the enzyme to use nicks and gaps in the DNA as starting points for labelling DNA by nick translation.

Advantages & Features
- Native: isolated from E. coli cells with a cloned fragment of the polA gene.
- DNase I-dependent nick translation: second-strand synthesis in cDNA cloning, fill-in of 5´ overhangs.
- Complete solution: supplied with 10x Reaction Buffer.

Specifications
Assay Conditions:
67 mM potassium phosphate (pH 7.4), 6.7 mM MgCl2, 1 mM 2-mercaptoethanol, 0.033 mMdATP, 0.033 mMdTTP, 0.4 MBq/mL [3H]-dTTP and 62.5 µg/mL poly(dA-dT)·poly(dA-dT).

Includes
- 50 μL DNA Polymerase I (10 U/μL)
- 1 mL Reaction Buffer (10x)

Applications
- High-percentage incorporation of radioactivity for nick translation assays.
- Manufacturing of alternating copolymers such as poly d(A-T) and homopolymers such as poly dG-poly dC.
- Klenow fragment: DNA sequencing, fill-in of 5’ overhangs and removal of 3’ overhangs to form blunt ends and second strand synthesis in mutagenesis.

Shipping & Storage
Shipped in: Gel pack.
Storage: -20 °C.