Klenow DNA Polymerase (3′ → 5′ exo-) for DNA Labeling

Klenow DNA Polymerase (3′ → 5′ exo-) is the large fragment of E. coli DNA polymerase I. It exhibits 5'→3' polymerase activity, but lacks the 3'→5' (proofreading) and 5'→3' (nick-translation) exonuclease activities of DNA Polymerase I. The 3'→5' exonuclease activity of the enzyme is eliminated by mutations in the 3'→5'-exonuclease active site (D355A and E357A ). Polymerase incorporates modified nucleotides (e.g., fluorescein-, biotin-labeled nucleotides).

Applications:

• Incorporation of fluorescent nucleotide analogs
• A-tailing and T-tailing of adapters for Next Generation Sequencing 
• Dideoxy DNA sequencing of single- or double-stranded DNA templates
• cDNA second-strand synthesis 
• Generate single-stranded DNA probes using random primers
• Site-directed DNA mutagenesis using synthetic oligonucleotides

Specifications:

Enzyme concentration: 5 Units/ul

Storage buffer: 20 mM Tris-HCl (pH 7.5), 0.1 mM EDTA, 1 mM DTT, and 50% (v/v) Glycerol

Optimal reaction temperature: 37 ^oC

Heat inactivation: 75 °C for 10 min

Storage: -20 ^oC

Shipping conditions: Shipped on dry ice

SKU: KLE-601-250

International Shipping:  Product requires shipping on dry ice. Please contact info@amidbiosciences.com for shipment estimates. 

Enzyme supplied with 10X Reaction Buffer (0.2 M Tris-HCl , pH 7.9, 0.5 M NaCl, 0.1 M MgCl₂ , 10 mM DTT)

Unit Definition: One unit is defined as the amount of Klenow DNA Polymerase (3′ → 5′ exo-) that catalyzes the incorporation of 10 nmol of dNTP into acid insoluble material in 30 minutes at 37°C under standard assay conditions.

This product is intended for laboratory research use only. Not for diagnostic purposes.