ArticleHorticulture research2021
An optimized protocol for stepwise optimization of real-time RT-PCR analysis.
Article in Horticulture research, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 59 papers.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
59 citing papers in PubMed, 122 citations in OpenAlex.
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- Identification of Reference Genes and Their Application in Analysis of Ginsenoside Biosynthesis Genes in American Ginseng Under Abiotic Stresses and Hormone Treatments.Plants (Basel, Switzerland) · 2026Article
- Validation of Optimal Reference Genes for qRT-PCR in Adipose- and Uterine-Derived Feline Mesenchymal Stem Cells.Veterinary sciences · 2026Article
- Wnt3a Downregulation Contributes to Type 2 Diabetes Mellitus by Impairing Pancreatic β-Cell Function.FASEB journal : official publication of the Federation of American Societies for Experimental Biology · 2026Article
- Transcriptomic Analysis Reveals HowLife (Basel, Switzerland) · 2026Article
- Multi-omics profiling reveals antagonistic strategies of endophytic Streptomyces sp. MEPP0209 against Lasiodiplodia theobromae causing Dendrobium officinale stem rot.Applied microbiology and biotechnology · 2026Article
- Chemerin 15 peptide reduces neuroinflammation via the ChemR23 receptor after ischemia-reperfusion injury.Neural regeneration research · 2026Article
- Reporting quality of quantitative polymerase chain reaction (qPCR) methods in scientific publications.Research integrity and peer review · 2026Article
- Bioactivity of green-synthesized zinc oxide nanoparticles using Fusarium oxysporum extract on the expression of extended-spectrum beta-lactamase and biofilm-associated genes in the pathogen Klebsiella pneumoniae.Journal, genetic engineering & biotechnology · 2026Article
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- Integrative analyses of metabolome and transcriptome reveal differences in terpenoids and identify the genes involved in the synthesis of terpenoids inFrontiers in plant science · 2026Article
- Optimization and validation of the real-time polymerase chain reaction protocol for porcine gelatin detection in processed food and cosmetic products.Open veterinary journal · 2026Article
- AoVps18 regulates sporulation, trap morphogenesis, and nematode predation by modulating vacuole assembly and attractant synthesis inVirulence · 2025Article
- Comparative transcriptome analysis reveals abscisic acid-induced bHLH transcription factors involved in saikosaponin biosynthesis inPlant signaling & behavior · 2025Article
- Viral Coinfections Potentially Associated with Feline Chronic Gingivostomatitis in Cats with Feline Infectious Peritonitis.Viruses · 2025Article
- [Preclinical Development Process and Prospects of Real-time Fluorescence Quantitative Polymerase Chain Reaction Detection Kits].Sichuan da xue xue bao. Yi xue ban = Journal of Sichuan University. Medical science edition · 2025Review
- Comparative Evaluation of Computational Methods for Validating Housekeeping Gene RT-qPCR Data in 3T3-L1 Cells.Biomedicines · 2025Article
- Let-7e as Potential Diagnostic Biomarkers in Early Detection of Peri-Implant Diseases: A Cross-Sectional Study.Clinical and experimental dental research · 2025Article
- Development of a SYBR Green-Based Real-Time PCR Assay to DetectTropical medicine and infectious disease · 2025Article
Corrections and comments
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Authors and funding
12 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Computational tool-assisted primer design for real-time reverse transcription (RT) PCR (qPCR) analysis largely ignores the sequence similarities between sequences of homologous genes in a plant genome. It can lead to false confidence in the quality of the designed primers, which sometimes results in skipping the optimization steps for qPCR. However, the optimization of qPCR parameters plays an essential role in the efficiency, specificity, and sensitivity of each gene's primers. Here, we proposed an optimized approach to sequentially optimizing primer sequences, annealing temperatures, primer concentrations, and cDNA concentration range for each reference (and target) gene. Our approach started with a sequence-specific primer design that should be based on the single-nucleotide polymorphisms (SNPs) present in all the homologous sequences for each of the reference (and target) genes under study. By combining the efficiency calibrated and standard curve methods with the 2
Identifiers
What OpenQuestion holds
Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.