ArticleBiomolecular detection and quantification2019
Shedding light: The importance of reverse transcription efficiency standards in data interpretation.
Article in Biomolecular detection and quantification, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed.
- A practical guide to targeted single-cell RNA sequencing technologies.Communications biology · 2026Review
- Scalable inference and identifiability of kinetic parameters for transcriptional bursting from single cell data.Bioinformatics (Oxford, England) · 2025Article
- Rapid and Sensitive Detection of RNA Viruses through Imaging of Marker Molecules Derived from Designed Circular DNA Probes.Small methods · 2025Article
- Mining, validating, and quantifying circular RNA transcriptome from total RNA as a biomarker or target.Scientific reports · 2025Article
- The Impact of the Variability of RT-qPCR Standard Curves on Reliable Viral Detection in Wastewater Surveillance.Microorganisms · 2025Article
- Establishment of genomic RNA reference materials for BCR-ABL1 P210 measurement.Analytical and bioanalytical chemistry · 2024Article
- Article
- Establishment of potential reference measurement procedure and reference materials for EML4-ALK fusion variants measurement.Scientific reports · 2024Article
- Identification and optimization of parameters for accurate quantification of RNA by RT-dPCR.Analytical and bioanalytical chemistry · 2024Article
- Exploring the impact of primer length on efficient gene detection via high-throughput sequencing.Nature communications · 2024Article
- Reducing Bias and Quantifying Uncertainty in Fluorescence Produced by PCR.Bulletin of mathematical biology · 2023Article
- Accurate quantification of SARS-CoV-2 RNA by isotope dilution mass spectrometry and providing a correction of reverse transcription efficiency in droplet digital PCR.Analytical and bioanalytical chemistry · 2022Article
- Statistics or biology: the zero-inflation controversy about scRNA-seq data.Genome biology · 2022Review
- Adaptive Immune Receptor Repertoire (AIRR) Community Guide to TR and IG Gene Annotation.Methods in molecular biology (Clifton, N.J.) · 2022Article
- STArS (STrain-Amplicon-Seq), a targeted nanopore sequencing workflow for SARS-CoV-2 diagnostics and genotyping.Biology methods & protocols · 2022Article
- Validation of the analytical performance of nine commercial RT-qPCR kits for SARS-CoV-2 detection using certified reference material.Journal of virological methods · 2021Article
- Tutorial: Guidelines for Single-Cell RT-qPCR.Cells · 2021Review
- Application of One-Step Reverse Transcription Droplet Digital PCR for Dengue Virus Detection and Quantification in Clinical Specimens.Diagnostics (Basel, Switzerland) · 2021Article
- A PCR assay to quantify patterns of HBV transcription.The Journal of general virology · 2021Article
- High Throughput Sequencing for the Detection and Characterization of RNA Viruses.Frontiers in microbiology · 2021Review
Corrections and comments
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Authors and funding
3 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The RNA-to-cDNA conversion step in transcriptomics experiments is widely recognised as inefficient and variable, casting doubt on the ability to do quantitative transcriptomics analyses. Multiple studies have focused on ways to optimise this process, resulting in contradictory recommendations. Here we explore the problem of reverse transcription efficiency using digital PCR and the RT method's impact on subsequent data analysis. Using synthetic RNA standards, an example experiment is presented, outlining a method to (1) determine relevant efficiency and variability values and then to (2) incorporate this information into downstream analyses as a way to improve the accuracy of quantitative transcriptomics experiments.
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