real-time PCR - How to design primers (Jun/07/2004 )

hi,

I am going to do real-time PCR using Smart Cycler, but I have never

done this experiment, I do not know how to design primers,such as the

length of primers,the size of PCR product and the annealing temperature

et al.

If you happen to know the knowledge about it, would you like to

sharing with me?

Thanks a lot!

Here you can find SmartCycler specific information on primer design for real-time PCR.

http://biology.queensu.ca/~tyshenko/settingup.htm

Additional guidelines can be found here http://core.biotech.hawaii.edu/g-rtpcr.htm
If you are going to use SYBR Green I dye, primer design is similar to regular RT-PCR. However, since SYBR Green I dye will detect all double-stranded DNA, including non-specific reaction products, optimization of the reaction conditions for each primer set are very important.

SAGE

Thank you for your kindly reply !

Best wishes to you!