I am currently validating 18S rRNA, HPRT1 and GAPDH as housekeeping genes to normalize data obtained from my case:control qPCR study using human peripheral blood.
Group 1 - Patients (cases) with significant stenosis in any of the 3 coronary arteries (angiographically-verified)
Group 2 - Controls with no or
Good day,
I don't really understand how your samples are categorized, for a better answer I'd así you for more information about each group. However, 3 ct is not such a striking change, i mean, depending in the average transcript number, and standard deviation.
Adán Valenzuela Castillo Hi. Thanks for replying. So, my samples are classified as:
Group 1 - Patients (cases) with significant stenosis in any of the 3 coronary arteries (angiographically-verified)
Group 2 - Controls with no or
Alright, this could mean that your candidate HK genes are not stable expressed under your experimental conditions (SD > 1.5 Ct), OR that there could be some gDNA leftovers. My advise is to run you DNase digested RNA samples in agarose electrophoresis to see if there is any amount of gDNA. If you have any doubt about your results. Try running a PCR with your digested RNA samples as template, 18S would be a good one to further look for an amplification product. If gDNA is present, or mtDNA, you could be over estimating the gene expression data. Good luck.
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