ArticleBMC molecular biology2016
A reliable method for quantification of splice variants using RT-qPCR.
Article in BMC molecular biology, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 25 papers.
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Who cites it
25 citing papers in PubMed.
- RAGE-Mediated Signalling in Gynaecological Disorders: Review of Molecular Mechanisms and Therapeutic Perspectives.Expert reviews in molecular medicine · 2026Review
- The human branchpoint-interacting stem-loop sequence and structure regulates U2 snRNA expression, branchpoint recognition, and the transcriptome.Nucleic acids research · 2026Article
- Machine learning framework for mRNA alternative splicing analysis identifies a signature of progression in colorectal adenocarcinoma.Scientific reports · 2026Article
- Detection of ExpressedBiology · 2025Article
- The human branchpoint-interacting stem loop sequence and structure regulates U2 snRNA expression, branchpoint recognition, and transcriptome.bioRxiv : the preprint server for biology · 2025Article
- Characterization of aFrontiers in immunology · 2025Article
- Targeted Variant Assessments of Human Endogenous Retroviral Regions in Whole Genome Sequencing Data Reveal Retroviral Variants Associated with Papillary Thyroid Cancer.Microorganisms · 2024Article
- Investigating the NRAS 5' UTR as a target for small molecules.Cell chemical biology · 2023Article
- Transcriptome analyses in infertile men reveal germ cell-specific expression and splicing patterns.Life science alliance · 2023Article
- Structural, topological, and functional characterization of transmembrane proteins TMEM213, 207, 116, 72 and 30B provides a potential link to ccRCC etiology.American journal of cancer research · 2023Article
- Transcriptomic complexity of the human malaria parasite Plasmodium falciparum revealed by long-read sequencing.PloS one · 2022Article
- Mebendazole Mediates Proteasomal Degradation of GLI Transcription Factors in Acute Myeloid Leukemia.International journal of molecular sciences · 2021Article
- Single well, single-common primer pair, dual probe, duplex qPCR assay for the quantification of mRNA splicing variants.Biology methods & protocols · 2021Article
- High expression of CIN85 promotes proliferation and invasion of human esophageal squamous cell carcinoma.Molecular medicine reports · 2021Article
- APOL1 C-Terminal Variants May Trigger Kidney Disease through Interference with APOL3 Control of Actomyosin.Cell reports · 2020Article
- Apoptosis in Cancer Cells Is Induced by Alternative Splicing of hnRNPA2/B1 Through Splicing of Bcl-x, a Mechanism that Can Be Stimulated by an Extract of the South African Medicinal Plant,Frontiers in oncology · 2020Article
- The RNA hairpin binder TRIM71 modulates alternative splicing by repressing MBNL1.Genes & development · 2019Article
- Alternative splicing of the Izumo1 gene ensures triggering gamete fusion in mice.Scientific reports · 2019Article
- Next generation sequencing reveals novel alterations in B-cell heavy chain receptor repertoires associated with acute-on-chronic liver failure.International journal of molecular medicine · 2019Article
- Real-time PCR quantification of spliced X-box binding protein 1 (XBP1) using a universal primer method.PloS one · 2019Article
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2 authors.
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Abstract
backgroundThe majority of protein isoforms arise from alternative splicing of the encoding primary RNA transcripts. To understand the significance of single splicing events, reliable techniques are needed to determine their incidence. However, existing methods are labour-intensive, error-prone or of limited use.
resultsHere, we present an improved method to determine the relative incidence of transcripts that arise from alternative splicing at a single site. Splice variants were quantified within a single sample using one-step reverse transcription quantitative PCR. Amplification products obtained with variant specific primer pairs were compared to those obtained with primer pairs common to both variants. The identities of variant specific amplicons were simultaneously verified by melt curve analysis. Independent calculations of the relative incidence of each variant were performed. Since the relative incidences of variants have to add upto 100%, the method provides an internal control to monitor experimental errors and uniform reverse transcription. The reliability of the method was tested using mixtures of cDNA templates as well as RNA samples from different sources.
conclusionThe method described here, is easy to set up and does not need unrelated reference genes and time consuming, error-prone standard curves. It provides a reliable and precise technique to distinguish small differences of the relative incidence of two splice variants.
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