ArticleNucleic acids research2005
PCR inhibition by reverse transcriptase leads to an overestimation of amplification efficiency.
Article in Nucleic acids research, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 51 papers.
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Who cites it
51 citing papers in PubMed.
- Volumetric DNA microscopy for mapping spatial transcriptomes in three dimensions.Nature protocols · 2026Review
- Selection and Expression Stability Analysis of Housekeeping Genes for Real-Time PCR Normalization in Rhesus Macaque Peripheral Blood.Animals : an open access journal from MDPI · 2026Article
- An Optimized RT-qPCR Protocol for Comprehensive Analysis of microRNAs and mRNAs inNon-coding RNA · 2026Article
- Subtype-Dependent Expression Patterns of Core Hippo Pathway Components in Thymic Epithelial Tumors (TETs): An RT-qPCR Study.Biomedicines · 2026Article
- Analysis of qPCR Data: From PCR Efficiency to Absolute Target Quantity.International journal of molecular sciences · 2025Review
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- Lyophilised Reverse Transcriptase and Polymerase for Point-of-Care Diagnostics of SARS-CoV-2.Microbial biotechnology · 2025Article
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- Comparison of Commercially Available Thermostable DNA Polymerases with Reverse Transcriptase Activity in Coupled Reverse Transcription Polymerase Chain Reaction Assays.Methods and protocols · 2025Article
- Variation of gene ratios in mock communities constructed with purified 16S rRNA during processing.Scientific reports · 2024Article
- Analysis ofBiomedicines · 2024Article
- Biporous silica nanostructure-induced nanovortex in microfluidics for nucleic acid enrichment, isolation, and PCR-free detection.Nature communications · 2024Article
- Differential expression of Dusp1 and immediate early response genes in the hippocampus of rats, subjected to forced swim test.Scientific reports · 2023Article
- Analysis ofBioMed research international · 2023Article
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- Identification of novel genes involved in the biofilm formation process of Avian Pathogenic Escherichia coli (APEC).PloS one · 2022Article
- Tutorial: Guidelines for Single-Cell RT-qPCR.Cells · 2021Review
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- Suggested guidelines for validation of real-time PCR assays in veterinary diagnostic laboratories.Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc · 2020Article
- Expression analysis of genes involved in mitochondrial biogenesis in mice with MPTP-induced model of Parkinson's disease.Molecular genetics and metabolism reports · 2020Article
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Authors and funding
2 authors.
Funding
Abstract
This study addresses the problem of PCR inhibition by reverse transcriptase. It has been shown that the inhibition occurs mostly when a small amount of RNA is taken for RT reaction, and it is more visible for rarely expressed transcripts. We show here that the inhibition takes place regardless of what amount of template is utilized for RT. The inhibition possesses a global nature, i.e. the amplification of any given transcript may be compromised with different levels of inhibition. The process of inhibition also explains wrongfully derived PCR amplification efficiencies, sometimes more than 100%, when the sequential dilutions of unpurified RT sample are utilized to build the calibration curve. The RT influences PCR not only by inhibiting it. When microgram(s) of RNA are taken for RT reaction, reverse transcriptase may cause overamplification of some transcripts under certain PCR conditions. The possible mechanism of RT influence on PCR is presented, and a purification method is implemented to remove the effects of RT on PCR.
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