Shedding light: The importance of reverse transcription efficiency standards in data interpretation
| Autor: | Jessica L. Schwaber, Stacey B. Andersen, Lars K. Nielsen |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Process (engineering)
Computer science lcsh:QR1-502 Efficiency 01 natural sciences Biochemistry Modelling lcsh:Microbiology cDNA complementary DNA Quantitative PCR 03 medical and health sciences Structural Biology Digital polymerase chain reaction Original Research Article dPCR digital PCR Transcriptomics Molecular Biology lcsh:QH301-705.5 030304 developmental biology 0303 health sciences RT reverse transcription 010401 analytical chemistry Data interpretation Reverse transcription Complementary DNA Reverse transcriptase 0104 chemical sciences lcsh:Biology (General) Molecular Medicine Biochemical engineering qPCR quantitative PCR Digital PCR |
| Zdroj: | Biomolecular Detection and Quantification, Vol 17, Iss, Pp-(2019) Biomolecular Detection and Quantification |
| ISSN: | 2214-7535 |
| Popis: | 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. Keywords: Transcriptomics, Quantitative PCR, Digital PCR, Reverse transcription, Efficiency, Complementary DNA, Modelling |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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