Greetings,
We are conducting an experiment after which we will measure the levels of certain microRNAs in the hippocampi of albino rats.
What would be the best housekeeping/reference gene to use for normalization?
Also, Taqman system for qRT-PCR is to be used, which universal master mix would be better: with UNG or without?
Thank you.
Hi, Dr nada
i suggest you read these:
1- Basal microRNA expression patterns in reward circuitry of selectively bred high-responder and low-responder rats vary by brain region and genotype
Article Basal microRNA expression patterns in reward circuitry of se...
2- Normalization of microRNA expression levels in quantitative RT-PCR assays: Identification of suitable reference RNA targets in normal and cancerous human solid tissues
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2327352/
3- Which is the accurate data normalization strategy for microRNA quantification?
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4890630/
good luck
Boris Schmitz
Although miR-191, miR-103, and miR-17–5p are good choices, but as we have used U6 in our experiments for leukemia cell lines and got good results I think U6 also works very well. It is also mentioned in some articles:
https://www.nature.com/articles/s41598-018-29297-7
I would suggest to use endogenous molecules, such as the one mentioned above. But better to check the inter-samples variability of the normalizator before trusting it faithfully.
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