relative quantification - (Aug/09/2006 )
Hello
I am new to relative quantification using RT-PCR. I dont know how to quantify cDNA for the assay. Now i am taking 5ug of RNA to convert to cDNA, from the cDNA iam taking 10 ul for all the samples is this correct? it will of great useful if any of you suggest on this experiment.
Devi
because the quantification of cDNA is problematic, many people quantify their RNA and add as close as possible to the same amount to every tube...just as you are doing.
you may need to change the amount of template you add when you are doing your optimization (if you haven't done it yet?) but in my opinion you are starting off correctly and your input amount is always relative to the amount of RNA added.
you may need to change the amount of template you add when you are doing your optimization (if you haven't done it yet?) but in my opinion you are starting off correctly and your input amount is always relative to the amount of RNA added.
Hello
Thankyou very much for your advice.
Devi
To quantify your DNA use a Nanodrop. If your department doesn't have one see if any others do.