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Klenow Enzyme

ROCHE/10104531001 - Sequencing grade, from Escherichia coli Iysogenic for NM 964

Synonym: Klenow

Product Type: Chemical

Catalog Number PKG Qty. Price Quantity
45-10104531001 250 units
$0.00
1/EA
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biological source Escherichia coli
form solution
manufacturer/tradename Roche
mol wt Mr 75 kDa
packaging pkg of 250 U (5 x 103 U/ml)
Quality Level 100 
specific activity >5000 units/mg protein
storage temp. −20°C (−15°C to −25°C)
Application: Klenow Enzyme has been used in blunt end ligation of recyclin-1 (RCY1) gene into pETDUET-GSTHis6-S Tag vector.
General description: Klenow enzyme is the large, C-terminal fragment (Mr 76,000) of E.coli DNA polymerase I, which can be obtained by subtilisin treatment of intact DNA polymerase I. It retains the 5′→3′ polymerase and the 3′→5′ exonuclease activities of intact DNA polymerase I, but lacks the 5′→3′ exonuclease activity of the native enzyme. The enzyme catalyzes the addition of mononucleotides from deoxynucleoside-5′-triphosphates to the 3′-hydroxyl terminus of a primer/template DNA. This property is used to synthesize DNA complementary to single-stranded DNA templates.
Klenow Enzyme is formed by subtilisin treatment of DNA polymerase I.
Other Notes: For life science research only. Not for use in diagnostic procedures.
Physical form: Solution of enzyme, 5U/μl, in storage buffer:
50 mM Potassium phosphate, 1 mM dithioerythritol, glycerol, 50% (v/v); pH 7.0 (4 °C)
Quality: Function test: Sequencing Grade Klenow Enzyme is tested in sequencing reactions according to Sanger. The test is conducted with 1 unit of enzyme and the M13 sequencing system. Sequencing gels are examined by autoradiography. The readable sequence must be ≥250 bases.

Absence of contaminants: Sequencing Grade Klenow Enzyme is tested on various DNA substrates to ensure the absence of nonspecific endonucleases and exonucleases.
Unit Definition: One unit is the enzyme activity which incorporates 10 nmol of total nucleotides into an acid-precipitable fraction in 30 minutes under the following assay conditions:
Incubation Buffer
130 mM Potassium phosphate, 6.5 mM MgCI2, 33 μM 14C-dTTP, poly[d(A-T)], 0.833 A260/ml; dATP, 33 μM; 1 mM dithioerythritol; bovine serum albumin, 0.032 mg/ml; pH 7.4.
Incubation Procedure
0.05 to 0.25 units of enzyme are incubated for 30 minutes at +37 °C in a total volume of 300 μl incubation buffer.

Volume Activity: 1,000 units/ml, or 5,000 units/ml respectively, according to Richardson (+37 °C, poly [d(A-T)] as primer).
RIDADR NONH for all modes of transport
WGK Germany WGK 1
Flash Point(F) does not flash
Flash Point(C) does not flash
activity specific activity: >5000 units/mg protein
Storage Temp. −20°C (−15°C to −25°C)
UNSPSC 41106314

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