Sybr green RT-qPCR primer - (Jun/14/2011 )
hi,i am a newbie in the RT-qPCR. I will on study the gene expression of different interluekin genes by SYBRgreen RT-qPCR. I dont want to design the primers myself and i have found journals who have the primer sequence for those interleukin genes. However, these primer sequences are designed for taqman RT-qPCR analysis. Here, i would like to ask can i use those primer sequences (without taqman probe) and subjected the primers to sybr green RT-qPCR?
Besides that, one step RT-qPCR or two-step RT-qPCR is of better choice if i want to study lots of interlukine genes(around 15 genes) from a tissue sample?
Finally, can someone share with me on the flow of running the RT-qPCR? (A simple one will help me lots as i have read lots journal and i actually end up with confious)
You can use them without probe, but check the specificity, probe-based assays are extra specific. Also check the formation of dimers of that primers.
What exactly are you asking about? What to do first when your primers arrive? How to optimise the reaction? How to pipette it? How to prepare lab for doing qPCR?