excess amount of primers - (Jun/10/2014 )
has anyone ever had problems with an excess amount of primers during the sequencing? I, by accident, added twice the amount of primers than supposed to.
Would this really matter this much? I can hardly imagine what the primers would do to the reaction to stop it ?
I know that if I wait, I'll know whether it worked or not, but I am just curious what could go wrong with the primers? More chances of primer dimers if the amount is too much?
It will likely work fine.
since you only use one primer when sequencing there is little chance of seeing primer dimers (unless they are self annealing).
too much primer may cause a burst of shorter sequences then a rapid drop in later fragments as they use up the terminator (dideoxy) nucleotides. but this is more likely to happen if you have too much template, as well.