difference between hotstart and taq polymerase - (Jan/26/2009 )

Hi everyboby,

what is the difference and the purpose of using hot start polymerase over Taq polymerase for pcr amplification

thanx

hot start enzymes are activated at high temps, so you can set up your reactions at room temp. Usually hot start enzymes are more accurate than convinient Taq enzymes.

hi
does hotstart help in reducing the non specific amplifications.
thanx a lot

It is said to do and in some of my experiments it did.

But if you have the money: I would try phusion (Finnzymes) or Accuprime / Amplitaq (Invitrogen). I had best results with those! But I am working with environmential samples, maybe there is some better enzyme round for your application.

Hi
At what temperature and how much time does it require to be activated.
thanx

novagen on Jan 26 2009, 11:36 AM said:

info comes with the enzyme but i believe that it activates during the normal denaturing step at the start of the pcr.

95degC for between 5-15mins

depends...my Qiagen one needs 95 degree C for 15 mins and it works pretty well.