KlenThermPlatinum™ DNA polymerase is a modified form of KlenThermTM DNA polymerase, that offers excel-lent 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 mis-matches which results in a very low rate of mis-match extension.
KlenThermPlatinum™ 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 mis-match 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 inconvenient 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.
- PCR requiring high specificity
- PCR with GC-rich regions or repeats (e.g. microsatellites)
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 MgCl 2 , 1 mM .-mercaptoethanol) and activated calf thymus DNA as substrate.
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)
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!
- 1.5 ml 10x reaction buffer Cat. No GC-001-006
Activity, non-specific endonucleases/nickases and exonucleases.