Greetings,

The most important dispersion measure to estimate the suitability ir your HK genes is the standard deviation. I'm not sure about what am I seeing at the attaches imagen. However, STD deviation among your samples should be less than 1 Ct. There are several methods to perform the estimation of the expression stability, you can go with geNorm, NormFinder and other classical freely available software. If you wish, look at my profile for the paper we published in 2015, where 4 distinct protocols were used to evaluate the suitability of 4 HK genes, yo final run pearson's correlation to discard any kind of bias due to each method.

Good luck

Hi Hafiz Muhammad Rizwan ,

Some tips for you:

1) The CV% i accept is 25%. There is no golden rule for this. The CV% must be calculated after 2-Cq transformation. Do not calculate CV% with Cq (Ct) since real time data has a exponential nature.

2) To choose the amplification range with best repeatibility, maintain your samples and standard curve between 15-25 cycles. Do not accept amplification beyond 30 cycles. I recommend this article to full information about real time pcr optimization. Article The Ultimate qPCR Experiment: Producing Publication Quality,...

Best,

Jacó

Adán Valenzuela Castillo please send PDF of the paper you published in 2015. Thank you in advance.

@Francis Nunes, of course, just send me your email address vía private message and I'll get back at you as soon as I can.

Sorry I can't find how to send it via private message, Adán Valenzuela Castillo.

Here we go: [email protected]

Thanks a lot.

Francis Nunes, I sent you the paper already. Should you have any question, please let me know.

I just received it, thanks a lot!

Adán Valenzuela Castillo Thanks a lot for your nice answer, Can you please share with me your 2015 article

Here is my email: [email protected]

Thanks

Hi there Hafiz, I'm sorry, I meant 2017 MS. Anyhow, I've just sent it, so it should be on your inbox already. Good Luck.