qPCR primer design tutorial

[mooseo]

Hi,
I'm fairly new to qPCR and would love to find a good tutorial on primer design. I'm currently using Invitrogen Vector NTI software, but I've tried other tools and they all see to spit out a bunch of likely looking options.

Specifically, how do I decide on the most important design considerations? Should I be especially concerned about Tm or are primer dimers and hairpins a bigger worry? I've received lots of helpful advice from around the lab, but it seems like a mash of rules of thumb (avoid 4 A in a row).

If anyone knows of a good site, or has a list of rules written down, that would certainly help me.

Thanks,
mike

[danfive]

Look at OLIGO software and website. The rule of thumb you mention about the 4As in a row is quite serious 4As are very weak bonds that will create a weak area of very low deltaG, these simply will not work 99% of the time for primers.