RT-qPCR normalizationRT-qPCR normalization
Posted 30 October 2012 - 10:38 AM
I'm new in RT-qPCR.
So I isolated my RNA and did a Nanodrop Spec on the RNA ng/uL. Then I did reverse transcription using Taqman box. Our lab usually do 500 ng RNA per reaction (18uL). But one of my RNA samples only had ~32 ng/uL, so I didn't dilute it. Now my final RNA/rxn for this sample is ~303 ng, while other samples are all ~500ng.
Then I did qPCR on the cDNA I got. I plotted the relative abundance.
Is there anyway to normalize the qPCR data because I started with different amount of RNA during RT? Or is the relative abundance already normalize the difference for me?
And what is relative expression level?
Posted 06 November 2012 - 08:44 PM
Posted 08 November 2012 - 10:33 PM
Posted 15 November 2012 - 05:07 AM