7 replies to this topic

[novagen]

Hi everyboby,

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

thanx

[gebirgsziege]

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.

[novagen]

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

[gebirgsziege]

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.

[novagen]

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

[mdfenko]

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

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

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

[carlchurchill]

95degC for between 5-15mins

[Lazinase]

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