ArticleFrontiers in genetics2022
Improved Nanopore full-length cDNA sequencing by PCR-suppression.
Article in Frontiers in genetics, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed.
- Review
- Distinct 5' and 3' coverage biases shape transcriptome interpretation in Nanopore direct RNA versus PCR-cDNA sequencing.BMC genomics · 2026Article
- Sensitive, flexible, and affordable serum RNA sequencing for pathogen detection on the Oxford Nanopore platform.BMC genomics · 2026Article
- Enhanced detection of RNA modifications in Escherichia coli utilizing direct RNA sequencing.Cell reports methods · 2025Article
- Long-read RNA sequencing of archival tissues reveals novel genes and transcripts associated with clear cell renal cell carcinoma recurrence and immune evasion.Genome research · 2024Article
- Comparison of direct cDNA and PCR-cDNA Nanopore sequencing of RNA fromMicrobial genomics · 2024Article
- CapTrap-seq: a platform-agnostic and quantitative approach for high-fidelity full-length RNA sequencing.Nature communications · 2024Article
- An optimized workflow of full-length transcriptome sequencing for accurate fusion transcript identification.RNA biology · 2024Article
- An Efficient Method to Prepare Barcoded cDNA Libraries from Plant Callus for Long-Read Sequencing.Methods and protocols · 2023Article
Corrections and comments
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Full-length transcript sequencing remains a main goal of RNA sequencing. However, even the application of long-read sequencing technologies such as Oxford Nanopore Technologies still fail to yield full-length transcript sequencing for a significant portion of sequenced reads. Since these technologies can sequence reads that are far longer than the longest known processed transcripts, the lack of efficiency to obtain full-length transcripts from good quality RNAs stems from library preparation inefficiency rather than the presence of degraded RNA molecules. It has previously been shown that addition of inverted terminal repeats in cDNA during reverse transcription followed by single-primer PCR creates a PCR suppression effect that prevents amplification of short molecules thus enriching the library for longer transcripts. We adapted this method for Nanopore cDNA library preparation and show that not only is PCR efficiency increased but gene body coverage is dramatically improved. The results show that implementation of this simple strategy will result in better quality full-length RNA sequencing data and make full-length transcript sequencing possible for most of sequenced reads.
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