ArticleInternational journal of legal medicine2012
Stability of endogenous reference genes in postmortem human brains for normalization of quantitative real-time PCR data: comprehensive evaluation using geNorm, NormFinder, and BestKeeper.
Article in International journal of legal medicine, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 77 papers.
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77 citing papers in PubMed.
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- Carnosine modulates Aβ-induced transcriptional aberrations in murine microglial cells.Current research in pharmacology and drug discovery · 2025Article
- Reference Gene Selection for RT-qPCR Normalization inInternational journal of molecular sciences · 2024Article
- Forensic neuropathology in the past decade: a scoping literature review.Forensic science, medicine, and pathology · 2024Article
- Advancements in Reference Gene Selection for Fruit Trees: A Comprehensive Review.International journal of molecular sciences · 2024Review
- Natural infection of hybrid sturgeon (Frontiers in immunology · 2024Article
- Evaluation of Reference Genes for Normalizing RT-qPCR and Analysis of the Expression Patterns ofInternational journal of molecular sciences · 2023Article
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- Development of reference genes for RT-qPCR analysis of gene expression in Pleurotus pulmonarius for biotechnological applications.Scientific reports · 2023Article
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- A Strategy for the Selection of RT-qPCR Reference Genes Based on Publicly Available Transcriptomic Datasets.Biomedicines · 2023Article
- Mfsd2a attenuated hypoxic-ischemic brain damage via protection of the blood-brain barrier in mfat-1 transgenic mice.Cellular and molecular life sciences : CMLS · 2023Article
- Screening and Verification of Reference Genes for Analysis of Gene Expression in Garlic (Plants (Basel, Switzerland) · 2023Article
- Effect of Savirin or Ticagrelor Treatment on the Expression of Commonly Used Reference Genes inMicroorganisms · 2023Article
- Identification of RT-qPCR reference genes suitable for gene function studies in the pitaya canker disease pathogen Neoscytalidium dimidiatum.Scientific reports · 2022Article
- Evaluation of Reference Genes for Real-Time Quantitative PCR Analysis in Tissues from Bumble Bees (International journal of molecular sciences · 2022Article
17 more citing papers are in PubMed but not listed here.
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6 authors.
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Abstract
In forensic molecular pathology, quantitative real-time polymerase chain reaction (RT-qPCR) provides a rapid and sensitive method to investigate functional changes in the death process. Accurate and reliable relative RT-qPCR requires ideal amplification efficiencies of target and reference genes. However, the amplification efficiency, changing during PCR, may be overestimated by the traditional standard curve method. No single gene meets the criteria of an ideal endogenous reference. Therefore, it is necessary to select suitable reference genes for specific requirements. The present study evaluated 32 potential reference genes in the human brain of 15 forensic autopsy cases using three different statistical algorithms, geNorm, NormFinder, and BestKeeper. On RT-qPCR data analyses using a completely objective and noise-resistant algorithm (Real-time PCR Miner), 24 genes met standard efficiency criteria. Validation of their stability and suitability as reference genes using geNorm suggested IPO8 and POLR2A as the most stable ones, and NormFinder indicated that IPO8 and POP4 had the highest expression stabilities, while BestKeeper highlighted ABL1 and ELF1 as reference genes with the least overall variation. Combining these three algorithms suggested the genes IPO8, POLR2A, and PES1 as stable endogenous references in RT-qPCR analysis of human brain samples, with YWHAZ, PPIA, HPRT1, and TBP being the least stable ones. These findings are inconsistent with those of previous studies. Moreover, the relative stability of target and reference genes remains unknown. These observations suggest that suitable reference genes should be selected on the basis of specific requirements, experiment conditions, and the characteristics of target genes in practical applications.
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