i´m trying to amplify a 2kb and 548bp band from gDNA of mice but never got to see the big band. i don´t understand what happened! i´ve tried different conditions even to the point of putting 2 minutes for the extension but nothing happened.can anyone explain to me what had just occured?
thanks.
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drcliquet
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Posted 16 January 2004 - 06:59 AM
For me PCR is always a question of "magic".
I've tryed to set up the right conditions for many different amplifications. When the PCR is "good", it works in many different conditions.
But when I get no result, in my experience, changing parameters is just a waste of time.
The best thing to do is design a new set of primers and start form zero!
I've tryed to set up the right conditions for many different amplifications. When the PCR is "good", it works in many different conditions.
But when I get no result, in my experience, changing parameters is just a waste of time.
The best thing to do is design a new set of primers and start form zero!
