In gene expression studies, what would be the reliability of house keeping genes expression say in a disease/insect attacked tissue that is being subjected to studies (RT qPCR ). Will the comparison be proper or inconclusive? We expect the internal environment of the tissue to be disturbed and hence this question. Or, am I missing some thing which is already taken care of?
Excellent and alerting question..I think this need more exploration to confirm that.
Regards
yes, thats why it is recommended to use as much HK genes as you can and choose the most stable ones.
Thanks Igor Bychkov for the answer. Can you point to me a paper where such a study on what house keeping genes are to be selected, has been done?
Rajesh S Patil
There are kits like https://www.qiagen.com/sg/resources/resourcedetail?id=34d2f8ed-e8af-4c2a-a2af-ba9ed0f02632&lang=en (or thermo analog), that contains about 12 HK genes. For every specific experiment, you should test them and choose the most stable ones. You could synthesize your own qPCR primers for these genes, or choose HK genes from the most stable ones (e.g. based on Fantom5 expression data). I ussualy use EMC4, EIF3M, PSMA1, C11orf58, because they were much more stable in my specific experiment, when i compared them with more popular like B2M, HPRT, TFRC, TBP, GADPH, ACTB.
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