Taqman for 18s and SYBR for gene of intest.... does it make sense..

[widu]

Hi,
I am a newbie for qPCR. I am expected to run several real time experiments, but i have been asked to run a Taqman assay for the 18S gene (endogenous control) and SYBR green for the gene of interest.

For me it does not make sense as I will be trying to compare two chemistries... Am I wrong??? Please help...

widu

[amberpdidit]

I'd say you're totally right.

It is probably doable (since it is relative expression then as long as you are doing the same thing it will probably work out) but it is sort of ridiculous.

Why not just order some SYBR 18S primers (~10bucks and this is a good place to find a reliable one medgen.ugent.be/rtprimerdb/index.php)
and run both SYBR reactions. Your boss should love this because SYBR chemistry is WAY cheaper than TaqMan

[widu]

thanks amberpdidit.

[cmartino]

SYBR can be problematic because it is an intercolating dye that can give you a false signal (not as specific as a Taqman probe). If the gene of interest is available as a Taqman probe I would use that. Of course, if you're trying to pick up multiple variations of this target gene, that might be the motivation for going with SYBR. I'd be curious to know why you've been asked to use SYBR.

[admin]

thanks cmartino thanks a good point