ncRNAseq: simple modifications to RNA-seq library preparation allow recovery and analysis of mid-sized non-coding RNAs.
Future Science Ltd
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Minshall, N., Chernukhin, I., Carroll, J., & Git, A. (2022). ncRNAseq: simple modifications to RNA-seq library preparation allow recovery and analysis of mid-sized non-coding RNAs.. Biotechniques, 72 (1), 21-28. https://doi.org/10.2144/btn-2021-0035
Despite their abundance, mid-sized RNAs (30-300 nt) have not been extensively studied by high-throughput sequencing, mostly due to selective loss in library preparation. The authors propose simple and inexpensive modifications to the Illumina TruSeq protocol (ncRNAseq), allowing the capture and sequencing of mid-sized non-coding RNAs without detriment to the coverage of coding mRNAs. This protocol is coupled with a two-step alignment: a pre-alignment to a curated non-coding genome, passing only the non-mapping reads to a standard genomic alignment. ncRNAseq correctly assigns the highest read-numbers to established abundant non-coding RNAs and correctly identifies cytosolic and nuclear enrichment of known non-coding RNAs in two cell lines.
Reports, 5′ coverage, high-throughput sequencing, next-generation sequencing, non-coding RNA, size-selection
Breast Cancer Now (2016NovPR816)
External DOI: https://doi.org/10.2144/btn-2021-0035
This record's URL: https://www.repository.cam.ac.uk/handle/1810/333050