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difference between hotstart and taq polymerase


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7 replies to this topic

#1 novagen

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Posted 26 January 2009 - 06:55 AM

Hi everyboby,

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

thanx :(
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#2 gebirgsziege

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Posted 26 January 2009 - 06:57 AM

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.
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#3 novagen

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Posted 26 January 2009 - 07:11 AM

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

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Posted 26 January 2009 - 07:24 AM

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.
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#5 novagen

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Posted 26 January 2009 - 07:36 AM

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

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Posted 26 January 2009 - 08:15 AM

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.
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#7 carlchurchill

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Posted 07 May 2009 - 06:01 AM

95degC for between 5-15mins

#8 Lazinase

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Posted 07 May 2009 - 09:56 PM

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




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