Can I use genomic DNA to make a standard curve for qPCR
Posted 17 December 2009 - 08:42 PM
Posted 21 December 2009 - 10:01 AM
No you can't use the genomic DNA to get a standard curve. You will get false positive results as I have tried once. The other option I used was 18s rRNA as a control as well to quantify my qPCR. But since I used for the mammalian cells you can try to look out for the eukaryotic equivalent of 18s rRNA.
Posted 22 December 2009 - 10:09 AM
I thought that since genomic DNA contains all genes I want, it should be ok for me to use it as the template to make the standard curve. Is it right?
As for the internal control, I do use 16S rRNA.
Posted 23 December 2009 - 06:47 AM
an exception is an absolute standard curve if you are quantifiying mRNA, then you should also use reverse transcribed RNA (eg. IVT RNA) for your standard curve.
Posted 23 December 2009 - 09:15 AM
Posted 23 December 2009 - 08:15 PM
And also, unless you are very sure that the gene you want to amplify has only ONE copy per genome., Otherwise it is not a good idea to use genomic DNA.