Best enzyme for mutation analysis   High specificity   High signal/noise ratio   Difficult templates

DESCRIPTION
KlenThermPlatinum™ DNA polymerase is a modified form of KlenTherm™DNA polymerase, that offers excellent specificity. It is designed for PCR with difficult templates such as GC-rich fragments and microsatellites. KlenThermPlatinum™ is particularly well suited to primer extension of Single Nucleotide Polymorphism (SNP) markers.
KlenThermPlatinum™ maintains excellent specificity and minimal background even in conditions designed for high yield (high Mg²⁺ /primer concentrations). In fact, even on genomic templates, the enzyme can be used with MgCl²⁺ concentrations as high as 10 mM. KlenThermPlatinum™ has an extrem low signal/noise ratio. In addition, it has an extremely high recognition of base mismatches which results in a very low rate of mismatch extension. This enzyme is capable of extending through difficult regions, e.g. regions, which include inverted tandem repeats and those with high amounts of secondary structure.
KlenThermPlatinum™ works in a totally unique way, involving improved nucleotide selection at the active site, and a much lower rate of mismatch extension, meaning that only perfectly aligned primers will be extended. As a result, the enzyme can give even higher specificity than hot-start (manual or automatic) techniques without the need for pre-incubation steps. KlenThermPlatinum™ has a very weak terminal transferase activity, and products can be assumed to be blunt-ended. However, this is sequence dependent, and some sequences may be tailed with a single nucleotide.

APPLICATION
PCR requiring high specificity
PCR with GC-rich regions or repeats (e.g. microsatellites)
Mutation analysis

CONCENTRATION
10 units/µl

UNIT DEFINITION
One unit is defined as the amount of enzyme, that incorporates 10 nmoles of dNTP into acid-insoluble form in 30 min at 73°C under the assay conditions (25 mM TAPS pH 9.3 at 25°C, 50 mM KCl, 2 mM MgCl₂, 1 mM β-mercaptoethanol) and activated calf thymus 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
-20°C

10X REACTION BUFER
500 mM KCl, 100 mM Tris-HCl (pH 9 at 25°C), 1% Triton X100
Extra solution: 50 mM MgCl₂, add MgCl₂ to a final concentration of 3.5 mM.
Please note the difference between KlenTherm™ and BioTherm™ reaction buffers.

QUALITY CONTROL
Activity, non-specific endonucleases/nickases and exonucleases.