Problem using evagreen (evagreen vs sybrgreen) - (Jun/18/2013 )
I would like to know if anyone have wierd results using
I am sure but as I heard that somebody had the same problem in our university,
I think that some qPCR machines are not compatible with evagreen.
we use this machine and works well with evagreen:
althogh it is not durable.
Do you use normal evagreen ou Sso Fast evagreen? What do you mean by "although it's not durable"?
we use Sso Fast evagreen.
but I am not sure which kinds of evagreen has been used by that person.
Are you doing just quantitative PCR for expression studies or so?
In that case using SYBR alternative is AFAIK pointless, you don't need higher sensitivity for getting a Ct value and for melting curve it's true Evagreen could have higher discrimination ability but that's not really needed anyway, if you use melting just for checking the specifity of your product. It won't give you any additional information.
More sensitive intercalator dyes are usefull for HRM, but they usually affect amplification more in this applications and often need quite an optimization to work satisfactory.