Hello,
I use Taq polymerase, Recombinant (Invitrogen, #10342020)
In the its protocol, 0.1ul Taq is needed for 1 rxn of 25ul. But it is hard to take 0.1ul. So I think that I dilute Taq in 10 times by 1xBuffer and use 1 ul for 1rxn.
I am concerned that Taq would lose the activity when they are diluted and stored in -20 degree at frozen state.
Do you think that it is okay that using Taq polymerase dilution and freezing for storage?
Hello, Sooyong,
Commercially, Taq polymerases may come in different stock solutions (different concentrations) based on the manufacturer. You need to make sure that you are using the right concentration in your PCR protocol.
Yes you should be freezing your polymerase to prevent the enzyme from being less efficient the next time you use it. The enzyme will not lose its activity even if you freeze it. You just have to allow it thaw before your next intended use.
Hope this helps.
Best,
Arizaldo
I'm agree with Paolo, you should prepare a master mix for more than one sample or tube so that you can calculate Taq amount on the basis of Unit and add aquired amount of Taq.
Best regards.(ul)
Hot start enzymes are more stable than normal enzymes but all normal enzymes are shipped and stored in different buffer than reaction buffer. They are stored in high glycerol containing buffer so that the enzyme mixture does not freeze as the glycerol acts as an antifreeze . .Taq is quite stable at4c for storage short term but freezing will start to inactivate it. If you are storing diluted taq I think you should do it in storage buffer. the recipe for this is usually given in the information sheet that comes with the enzyme
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