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Experiment plan for RT-qPCR internal reference search

endogenous referenc

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#1 Abimbola Badejo

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Posted 20 July 2012 - 12:09 AM

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
http://www.protocol-...rs/page__st__30.

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...............?

#2 tea-test

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Posted 20 July 2012 - 02:00 AM

hi, do you have the old excel-based geNORM software?

I have uploaded the handbook where every step is explained in detail.

Attached Files


tea-test: The artist formerly known as Ned Land

#3 Abimbola Badejo

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Posted 20 July 2012 - 06:27 PM

Wow! Thank you tea-test. The manual is a great help. I will let you know of my progress after digesting the manual.Posted Image

#4 coffeebreak

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Posted 24 July 2012 - 12:35 AM

HI!

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?

Thanks!




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