Amid Biosciences| Competent Cells and Protein Expression Vectors
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).
- 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
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
International Shipping: Product requires shipping on dry ice. Please contact firstname.lastname@example.org 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.