ArticleScientific reports2017
A simple method to assess group difference in RT-qPCR reference gene selection using GeNorm: The case of the placental sex.
Article in Scientific reports, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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Who cites it
19 citing papers in PubMed.
- Analysis of qRT-PCR Data to Identify the Most Stable Reference Gene Using gQuant.Bio-protocol · 2025Article
- Placental expression and methylation of angiogenic factors in assisted reproductive technology pregnancies from India.Epigenomics · 2025Article
- Reference genes to study the sex-biased expression of genes regulating Drosophila metabolism.Scientific reports · 2024Article
- Advanced breast cancer diagnosis: Multiplex RT-qPCR for precise typing and angiogenesis profiling.Biochemistry and biophysics reports · 2024Article
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- Multi-Organ Transcriptome Response of Lumpfish (Microorganisms · 2022Article
- Validation of Reference Genes in a Population of BlueberryPlants (Basel, Switzerland) · 2022Article
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- Selection of reliable reference genes for analysis of gene expression in the rat placenta.Molecular and cellular biochemistry · 2021Article
- An Ex Vivo and In Silico Study Providing Insights into the Interplay of Circulating miRNAs Level, Platelet Reactivity and Thrombin Generation: Looking beyond Traditional Pharmacogenetics.Journal of personalized medicine · 2021Article
- Downregulation of the let-7 family of microRNAs may promote insulin receptor/insulin-like growth factor signalling pathways in pancreatic ductal adenocarcinoma.Oncology letters · 2020Article
- Reference Genes for Expression Studies in Human CD8Scientific reports · 2020Article
- Selection and evaluation of appropriate reference genes for RT-qPCR based expression analysis in Candida tropicalis following azole treatment.Scientific reports · 2020Article
- The Acute Effects of 5 Fluorouracil on Skeletal Muscle Resident and Infiltrating Immune Cells in Mice.Frontiers in physiology · 2020Article
- Development of a Bioinformatics Framework for Identification and Validation of Genomic Biomarkers and Key Immunopathology Processes and Controllers in Infectious and Non-infectious Severe Inflammatory Response Syndrome.Frontiers in immunology · 2020Article
- Article
- Selection of Reference Genes for Normalization of Real-Time PCR Data in Calliptamus italicus (Orthoptera: Acrididae) Under Different Temperature Conditions.Journal of insect science (Online) · 2019Article
- Impact of 5 fluorouracil chemotherapy on gut inflammation, functional parameters, and gut microbiota.Brain, behavior, and immunity · 2019Article
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3 authors.
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Abstract
Normalization with proper reference genes is a crucial step in obtaining accurate mRNA expression levels in RT-qPCR experiments. GeNorm and NormFinder are two commonly used software packages that help in selecting the best reference genes, based on their expression stability. However, GeNorm does not take into account a group variable, such as sample sex, in its calculation. We demonstrate a simple calculation step to assess the variability of such parameters by multiplying the GeNorm M value with the difference of Cq values between groups. To test this, we used 28 reference gene candidates, to analyze 20 placental samples (10 of each sex), and by using HPRT1 (lower Cq values in male placentas (P = 0.017)), as a target gene. Our calculation demonstrates that the RPL30 - GAPDH reference gene combination is the better option to assess small placental sex differences in mRNA level, versus the selection obtained from GeNorm or NormFinder. The HPRT1 normalized mRNA expression level is different between placental sexes, using RPL30 and GAPDH as reference genes (P = 0.01), but not when using genes suggested by GeNorm or NormFinder. These results indicate that the proposed calculation is appropriate to assess small variations in mRNA expression between 2 groups.
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