Sybr green RT-qPCR primer (View forum version)



rugs

Posted 14 June 2011 - 07:28 AM

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)

Trof

Posted 16 June 2011 - 07:40 AM

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.

Two-step.

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?