Dear all, I am working on differential expression of 5 miRNAs in 3 different accessions of tomatoes named Solanum pimpinellifolium, Solanum lycopersicum, and Solanum habrochaites under abiotic stresses. I am seeking the best reference gene which can be used for Real-time analysis studies. Also, I would also like to know how many numbers of reference genes could be sufficient for this type of study. I would be glad if you could guide me on this matter.
Thank you so much for your time and consideration in advance.
Regards,
Houtan
hi,
Check the papers, they have described plant miRNA housekeeping genes.
Article New insights into tomato microRNAs
Article Selection and validation of reference genes for quantitative...
Also, check the answers provided in the following discussion. https://www.researchgate.net/post/Which_houses_keeping_gene_is_suitable_as_endogenous_control_for_plant_miRNAs_normalixation_in_RT-qPCR
Regards
Saurabh
Hi,
I would not recommend using actin (and/or GAPDH) as a housekeeping gene for miRNA experiments, since research has shown that GAPDH and β-actin are direct targets of miR-644a.
Source: Article Housekeeping Gene Selection Advisory: Glyceraldehyde-3-Phosp...
Kind regards,
Tarik
You need to validate internal reference genes for each particular experiment. See MIQE guidelines. You can use software such as geNORM or Bestkeeper to analyse data from potential candidate reference genes
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