ThermoKlen™-Modified T. aquaticus DNA polymerase lacking 5´-3´ exonuclease activity.
Description	ThermoKlen™ (EC 2.7.7.7) is a N-terminally truncated Thermus aquaticus DNA polymerase where the domain responsible for the 5´-3´ exonuclease activity is removed. This deletion does not distort the 3´-5´ DNA-polymerase activity. As a result of this deletion, the enzyme is more heat-stable and is more suitable for amplification of GC-rich regions and templates with extensive secondary structures. The enzyme has a reduced tendency to extend a mismatched 3´ oligonucleotide end that make it more suitable for ARMS analysis. The enzyme has 3´ terminal deoxynucleotidyl transferase activity which usually results in addition of single dATP to the duplex DNA. Optimal temperature is 70°C, half-life at 95°C is about 1 h.
Unit definition	One unit of the enzyme will incorporate 10 nmol of dNTPs into an acid-insoluble form in 30 min at 72°C under standard assay conditions using a DNA template.
Reaction buffer	B5 (x10, with Mg2+ ): 670 mM Tris-HCl, pH 9.1, 166 mM (NH4)2SO4, 35 mM MgCl2, 0.15% BSA.
Storage	Store at -20°C.
Applications	Real-time quantitative PCR with hybridization probes, mutation analysis, site-directed mutagenesis, ARMS analysis, DNA labeling, primer extension, sequencing.
Quality control	activity, SDS-PAGE purity, absence of endo- and exonucleases, specific performance tests.
References	1. Barnes W.M. (1992) Gene 112, 29-35. 2. Wilhelm, J. et al. (2001) BioTechniques 30, 1052-1062.
Catalog #	T5.4	T5.5
Pack size	1000 U	2500 U