Klenow enzyme is a DNA-dependent 5′→3′ polymerase with 3′→5′ exonuclease activity. It lacks the 5′→3′ exonuclease activity of the native enzyme. The addition of mononucleotides from dNTPs to the 3′-OH terminus of DNA is catalyzed. This property is used to synthesize DNA complementary to single-stranded DNA templates.
Heat inactivation: Add 2 μl 0.2 mM EDTA (pH 8.0) and/or heat to 65 °C for 10 minutes
Use Klenow Enzyme for:
- Random-primed DNA labeling using random oligonucleotides as primers for the incorporation of nonradioactively labeled and 32P-labeled nucleotides.
- Fill-in reaction for blunt-end formation of 3′-recessed (staggered) ends.
Absence of endo- and exonuclease activity tested according to the current Quality Control procedures
One unit is the enzyme activity which incorporates 10 nmol of total nucleotides into an acid-precipitable fraction in 30 minutes under assay conditions at +37 °C with poly [d(A-T)] as primer (Richardson, C.C. & Kornberg, A. (1994) J. Biol. Chem. 244, 2996).
Volume Activity: 2,000 U/ml
Enzyme in Storage buffer: 50mM potassium phosphate, 1mM dithioerythritol, 50% glycerol (v/v), pH 7.0 (+4°C).
For life science research only. Not for use in diagnostic procedures.