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Finding cDNA for making a standard curve for real-time RT-PCR


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#1 ScienceAnna

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Posted 17 December 2009 - 02:02 AM

Hi all!

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

#2 tea-test

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Posted 17 December 2009 - 09:43 AM

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.
tea-test: The artist formerly known as Ned Land




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