Experiment plan for RT-qPCR internal reference search - (Jul/20/2012 )
hello, I'm trying to choose an internal reference gene among a group of 7 "HKG" for a bacterial gene expression study.
I've read some journals and I've also gone through most of the ones here
My problem is the experiment itself. I'm really confused about the genes.
1. Do I set up my RT-PCR plate with only one target gene and use all the ref genes in separate experiments, after which I'll collate all my data and load on gNORM or
2. I should use only one target gene as the "reference gene" , then use all the reference gene candidates as the "target genes"?
I'm actually a novice in qPCR and I've been struggling with this for the past 3 weeks.
I also have 6 samples ( 3 environmental conditions on a control and a test strain)
In summary, can someone please explain to me what I have to do, I beg you please...............?
Wow! Thank you tea-test. The manual is a great help. I will let you know of my progress after digesting the manual.
I have been looking for the old version of geNorm for ages!! (the one for excel). Does somebody know if it is still possible to get it and how?