ArticleBiological procedures online2021
Relative versus absolute RNA quantification: a comparative analysis based on the example of endothelial expression of vasoactive receptors.
Article in Biological procedures online, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed.
- The good, the bad, and the stable: Reference genes for preclinical biodistribution studies.Molecular therapy. Advances · 2026Article
- Transcriptional investigation of immune gene expression dynamics in Rohu (Labeo rohita) in response to killed Flavobacterium columnare vaccine.Molecular biology reports · 2025Article
- Extra-oral expression of bitter taste receptors in pigs and their correlation with hepatic cytochrome P450 enzymes.Cell and tissue research · 2025Article
- Extracellular RNAs in Liquid Biopsy: Applications in MASLD and MASH Diagnosis and Monitoring.Diagnostics (Basel, Switzerland) · 2025Review
- P-selectin glycoprotein ligand-1 and cardiovascular diseases: from a general perspective to an HIV infection context.Frontiers in cardiovascular medicine · 2025Article
- Advanced breast cancer diagnosis: Multiplex RT-qPCR for precise typing and angiogenesis profiling.Biochemistry and biophysics reports · 2024Article
- Quantitative evaluation of endogenous reference genes for ddPCR under salt stress using a moderate halophile.Extremophiles : life under extreme conditions · 2023Article
- An intronic GAA repeat expansion in FGF14 causes the autosomal-dominant adult-onset ataxia SCA50/ATX-FGF14.American journal of human genetics · 2023Article
- Identification of a Maturation Plasma Cell Index through a Highly Sensitive Droplet Digital PCR Assay Gene Expression Signature Validation in Newly Diagnosed Multiple Myeloma Patients.International journal of molecular sciences · 2022Article
- Application of Droplet Digital PCR Technology in Muscular Dystrophies Research.International journal of molecular sciences · 2022Review
- miRNA-Mediated Priming of Macrophage M1 Differentiation Differs in Gram-Positive and Gram-Negative Settings.Genes · 2022Article
Corrections and comments
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Authors and funding
3 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundIn the present study, two distinct PCR methods were used for the quantification of genetic material and their results were compared: real-time-PCR (qPCR; relative quantification) and droplet digital PCR (ddPCR; absolute quantification). The comparison of the qPCR and the ddPCR was based on a stimulation approach of microvascular endothelial cells in which the effect of a pro-inflammatory milieu on the expression of vasoactive receptors was investigated.
resultsThere was consistency in directions of effects for the majority of genes tested. With regard to the indicated dimension of the effects, the overall picture was more differentiated. It was striking that deviations were more pronounced if the measured values were on the extreme edges of the dynamic range of the test procedures.
conclusionsTo obtain valid and reliable results, dilution series are recommended, which should be carried out initially. In case of ddPCR the number of copies per µl should be adjusted to the low three-digit range. With regard to qPCR it is essential that the stability and reliability of the reference genes used is guaranteed. Here, ddPCR offers the advantage that housekeeping genes are not required. Furthermore, an absolute quantification of the sample can be easily performed by means of ddPCR. Before using ddPCR, however, care should be taken to optimize the experimental conditions. Strict indications for this methodology should also be made with regard to economic and timing factors.
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