KlenThermN™ DNA Polymerase

provides specific PCR amplification and sequencing of difficult templates,
Produkt Artikel-Nr. Packungsgröße Preise
KlenThermN™ DNA Polymerase
GC-023-0100
Manufacturer part number:
Availability:
100 u
17,50 € excl tax
KlenThermN™ DNA Polymerase
GC-023-0250
Manufacturer part number:
Availability:
250 u
43,75 € excl tax
KlenThermN™ DNA Polymerase
GC-023-0500
Manufacturer part number:
Availability:
500 u
87,50 € excl tax
KlenThermN™ DNA Polymerase
GC-023-1000
Manufacturer part number:
Availability:
1000 u
175,00 € excl tax
KlenThermN™ DNA Polymerase
GC-023-5000
Manufacturer part number:
Availability:
5000 u
875,00 € excl tax

DESCRIPTION

Substitution of Asn for the conserved Ser543 in the thumb subdomain of the Taq DNA polymerase large fragment (KlenTherm™ DNA polymerase) prevents pausing during DNA synthesis and allows the enzyme to overcome template regions with a complex secondary structure. The mutant enzyme, KlenThermN™ DNA poly-merase (patent pending), provides specific PCR amplification and sequencing of difficult templates, e.g. those with a high GC content or complex secondary structure. Furthermore this substitution increases several times the efficiency of synthesis of long (over 2 kb) DNA molecules. The difference in the DNA synthesis efficiencies by the mutant and native enzymes increases with the increase in the DNA fragment length.


CONCENTRATION

10 units/µl


UNIT DEFINITION

One unit is defined as the amount of enzyme that incorporates 10 nmoles of dNTPs into acid-insoluble form in 30 min at 73°C under the assay conditions 25 mM TAPS (tris-(hydroxy-methyl)-methyl-amino-propanesul-fonic acid, sodium salt) pH 9.3 (at 25°C), 50 mM KCl, 3.5 mM MgCl2 , 1 mM .-mercaptoethanol) and activa-ted salmon sperm DNA as substrate.


STORAGE BUFFER

10 mM K-phosphate buffer pH 7.0, 100 mM NaCl, 0.5 mM EDTA, 1 mM DTT, 0.01% Tween 20; 50% glycerol (v/v)


STORAGE TEMPERATURE

Store KlenThermN™ DNA polymerase, preferably at -20°C, in a constant temperature freezer.


10X REACTION BUFER

500 mM KCl, 100 mM Tris-HCl (pH 9 at 25°C), 1% Triton X100
Extra solution: 50 mM MgCl 2 , add MgCl 2 to a final concentration of 3.5 mM.
Please note the difference between KlenThermTM and BioThermTM reaction buffers!
1.5 ml 10x reaction buffer Cat. No GC-001-006


COMPANION PRODUCTS

KlenTherm™ DNA polymerase, Synergy™ DNA polymerase, SynergyN™ DNA polymerase, dNTPs


REFERENCES

  1. Ignatov, K.B., Miroshnikov, A.I., Kramarov, V.M. (1998) FEBS Lett. 425, 249-250. Substitution of Asn for Ser543 in the large fragment of Taq DNA polymerase increases the efficiency of synthesis of long DNA molecules.

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