Artifacts and biases of the reverse transcription reaction in RNA sequencing.
RNA sequencing
bias
reverse transcription
Journal
RNA (New York, N.Y.)
ISSN: 1469-9001
Titre abrégé: RNA
Pays: United States
ID NLM: 9509184
Informations de publication
Date de publication:
07 2023
07 2023
Historique:
medline:
19
6
2023
pubmed:
30
3
2023
entrez:
29
3
2023
Statut:
ppublish
Résumé
RNA sequencing has spurred a significant number of research areas in recent years. Most protocols rely on synthesizing a more stable complementary DNA (cDNA) copy of the RNA molecule during the reverse transcription reaction. The resulting cDNA pool is often wrongfully assumed to be quantitatively and molecularly similar to the original RNA input. Sadly, biases and artifacts confound the resulting cDNA mixture. These issues are often overlooked or ignored in the literature by those that rely on the reverse transcription process. In this review, we confront the reader with intra- and intersample biases and artifacts caused by the reverse transcription reaction during RNA sequencing experiments. To fight the reader's despair, we also provide solutions to most issues and inform on good RNA sequencing practices. We hope the reader can use this review to their advantage, thereby contributing to scientifically sound RNA studies.
Identifiants
pubmed: 36990512
pii: rna.079623.123
doi: 10.1261/rna.079623.123
pmc: PMC10275267
doi:
Substances chimiques
DNA, Complementary
0
RNA
63231-63-0
Types de publication
Review
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
889-897Informations de copyright
© 2023 Verwilt et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.
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