Finding cDNA for making a standard curve for real-time RT-PCR - (Dec/17/2009 )
I am analysing human mRNA expression with realtime RT-PCR technique, i.e I convert my mRNA to cDNA and analyse with Applieds Taqman probes and primers.
I use the standard curve method.
Now however, I have a problem that I havent found mRNA with enough expression of human MMP8 to be able to create a standard curve.
Does anyone know if you can buy cDNA anywhere?
Or how else could I solve this problem?
I really want to stick with the standard curve method since I already have analysed the expression of a house keeping control gene in all my samples with the standard curve method.
Kind regards Anna
hi, yes you can purchase cDNA or RNA from the appropriate companies like stratagene, clontech, ambion,...
my tip: instead of cDNA purchase RNA and reverse transcribe this with the same RT reagents you are using.
Another possibility: look, if there is a cDNA clone of your GOI available, eg. from Open Biosystems, Invitrogen, Imagenes,...
sometimes the clone costs only ~70 euros and you don't need the full length sequece, just the part, where your primers are binding.