My PCR reaction is good when I work by smartaq enzyme BUT isn't good by pfu

  • Thread starter Thread starter sahar p
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sahar p

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polymerase why? hello every body
this is a puzzle for me,!!!!! Im confused since when I work by Smartaq polymerase, bands are so sharp and excellent but when I work by pfu in PCR reaction i don't detect any band in gel( at the same or different conditions and with different gradients)
please guide me
many thanks
 
The two enzymes have different reaction conditions for optimal results. When you switch enzymes you may have to re-optimize the reaction. You should try varying all of the following conditions: template concentrations, primer concentration, Mg++ concentration, and annealing temperature. You can do two variables at a time on a 96 well plate (or even more if you divide the plate in two).

I don't know off the top of my head but certain enzymes are better for small products and others for large products.

However, your best best is to stay with what works. If it ain't broke, don't fix it!
 
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