storage of the homemade Taq polymerase - troubles with taq storage (Feb/15/2005 )
We have trouble with the storage of the homemade taq.
We use the Bio Rex 70 Resin for its purification.
We have tried with different glicerol qualities and concentracions, autoclavated or not.
With 50% glycerol it looses activity and with 30% it gets frozen.
What can we do?.
ABI Taq storage buffer is 20 mM Tris-HCl, pH 8.0 @ 25°C, 100 mM KCl, 0.1 mM Na2EDTA, 1 mM DTT, 0.5% (v/v) Tween 20, 0.5% (v/v) Nonidet-P40 and 50% (v/v) glycerol.
Glycerol will start to decompose to ethylene aldehyde at its boiling point of 182°C, so it should not be autoclaved. You can filter sterrilize 80% glycerol and use that to prepare the buffer.
Clean Taq preps are actually stable at room temperature for 6 months, but the enzyme is kept in freezers to prevent potential contaminants (bacterial or fungal) from growing.