primer dimers - (Nov/12/2005 )
hi all,
i got nice bands after multiplex pcr but got intense primer dimers as well. For pure dtection whether the gene is positive or not, is it okay if i have primer dimers?
What does experts have to say when i present a picture of a gel having primer dimers in them?
thanks
I assume you are doing RT-PCR just to show a presence of the transcript.
If the 'nice bands' you metioned are nice enough, I would say it is an acceptable data.
But it's probably not very useful for making any quantitative arguments.
By the way, some polymerases are much superior than others in terms of multiplex PCR. If you use one of those fancy Taq polymerases, the primer-dimer might disappear. Check out the QIAGEN catalog, for example. They have some good stuff.
why dont u dilute your primers further and get a good gel, if u want to present your data well.
May be ur primer concentration is too high for that template.........check out
yes, i was thinking of lowering the primer concentration of the other too. Thank you for your insights, i will try to make it better.
Hi,
I would try to get rid of the dimers by making new primers. Lots of free programmes like primer3 are free and online and alow you to make primers that are compatible with the ones you already have.
also very good is oligo, but its not free:(
ken,