KLEN-RO

Roche

Klenow Enzyme

Labeling grade, from Escherichia coli lysogenic NM 964

Enzyme Commission number:

Quality Level

form

solution

specific activity

>5000 units/mg protein

packaging

pkg of 100 U (11008404001)
pkg of 500 U (11008412001)

manufacturer/tradename

Roche

shipped in

dry ice

storage temp.

−20°C

General description

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.

Specificity

Heat inactivation: Add 2 μl 0.2 mM EDTA (pH 8.0) and/or heat to 65 °C for 10 minutes

Application

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.

Quality

Absence of endo- and exonuclease activity tested according to the current Quality Control procedures

Unit Definition

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

Physical form

Enzyme in Storage buffer: 50mM potassium phosphate, 1mM dithioerythritol, 50% glycerol (v/v), pH 7.0 (+4°C).

Other Notes

For life science research only. Not for use in diagnostic procedures.

RIDADR

NONH for all modes of transport

Certificate of Analysis

Certificate of Origin

Protocols
Klenow Enzyme Protocol
Read More

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