ArticleProtoplasma2017
A spiking strategy facilitates housekeeping selection for RT-qPCR analysis under different biotic stresses in eggplant.
Article in Protoplasma, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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Who cites it
5 citing papers in PubMed.
- Article
- AInternational journal of molecular sciences · 2021Article
- Identification and validation of superior housekeeping gene(s) for qRT-PCR data normalization inPhysiology and molecular biology of plants : an international journal of functional plant biology · 2020Article
- Article
- Molecular dynamics of pre-germinative metabolism in primed eggplant (Horticulture research · 2020Article
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Authors and funding
7 authors.
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Abstract
Endogenous housekeeping genes are traditionally employed to normalize the expression of target genes in RT-qPCR studies. Assuming that a perfect housekeeping suitable for every condition does not exist, expression stability of the chosen reference gene should be evaluated at every new experiment. The housekeeping selection process reveals furthermore complicated and time-consuming when different conditions have to be compared in the same experimental dataset. As an alternative strategy, we spiked an external reference transcript (ERT) into all RNA samples of our dataset (eggplant roots subjected to different biotic stresses), and used it to normalize the expression levels of native candidate housekeeping. ERT expression resulted highly stable across all samples and enabled to indicate glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as the most stable endogenous housekeeping. This result was confirmed by the use of GeNorm, Normfinder, and BestKeeper algorithms. This method might be generally applied to expedite the selection process of the best reference gene.
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