First, i want to tell you sorry for my bad english
there is my problem:
So, i need to make Master Mix =)
what a differense from basic PCR? I mean...eeemmm...Do anybody speak Russion?
Eh.. Do you know any links for literature about Master Mix for Real-Time PCR?
Smth like: Total concentration for reagents (total volume 50 ml):
DNA ~ 100-400 ng per reaction
Tag ~ 1 unit/ml ~ 1 ml per reaction
Primer A, Primer B ~ 1 ml per reaction (0.2 pMol) 10pMol/ml in Stock
MgCl2 ~ 2-2.5 mM per reaction
dNTP ~ 400 mM per reaction
Do i need Rox for detect Luminescence? What concentration of Probe do i need for reaction?
Unfortunately all of protocols ive seen, autors just wrote "WasterMix" without detalisation.