IMMOLASE is a heat-activated form of a thermostable DNA polymerase, which is inactive at room temperature and provides improved specificity when compared to standard DNA polymerases. IMMOLASE can eliminate the presence of non-specifics such as primer-dimers and mis-primed products. The enzyme must be activated by heat treatment before primer extension is possible. Subsequently, the reaction can be handled according to the user's existing protocols for thermostable DNA polymerases.Such pre-treatment will give excellent specificity, but even higher specificity can be achieved by reducing the time of the preincubation. This allows the amount of enzyme available in the reaction to increase gradually over time and reduces the likelihood of there being, in the essential early stages of the reaction, an excess of enzyme over template, which is known to cause problems with specificity. Unit Definition: One unit is defined as the amount of enzyme that incorporates 10nmoles of dNTPs into acid-insoluble form in 30 minutes at 72 C under the following assay conditions: 25mM TAPS (tris-[hydroxymethyl]methyl-amino-propane-sulphonic acid, sodium salt), pH 9.3 at 25 C, 50mM KCI, 2mM MgCI2, 1mM beta-mercaptoethanol, 200uM of each dATP, dGTP, dTTP, 100uM dCTP (a mix of unlabelled and alha 32P-labelled) and 12.5ug activated salmon sperm DNA in a final volume of 50ul.
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