| Title |
Sensitive and accurate analysis of gene expression signatures enabled by oligonucleotide-labelled cDNA / |
| Authors |
Kapustina, Žana ; Medžiūnė, Justina ; Dubovskaja, Varvara ; Matjošaitis, Karolis ; Žeimytė, Simona ; Lubys, Arvydas |
| DOI |
10.1080/15476286.2022.2078093 |
| Full Text |
|
| Is Part of |
RNA biology.. Philadelphia : Taylor & Francis Inc.. 2022, vol. 19, iss. 1, p. 774-780.. ISSN 1547-6286. eISSN 1555-8584 |
| Keywords [eng] |
RNA-seq ; transcriptomics ; modified nucleotides ; library preparation ; 3'-end sequencing ; gene expression profiling |
| Abstract [eng] |
High-throughput RNA sequencing offers a comprehensive analysis of transcriptome complexity origi-nated from regulatory events, such as differential gene expression, alternative polyadenylation and others, and allows the increase in diagnostic capacity and precision. For gene expression profiling applications that do not specifically require information on alternative splicing events, the mRNA 3′ termini counting approach is a cost-effective alternative to whole transcriptome sequencing. Here, we report MTAS-seq (mRNA sequencing via terminator-assisted synthesis) – a novel RNA-seq library pre-paration method directed towards mRNA 3′ termini. We demonstrate the specific enrichment for 3′- terminal regions by simple and quick single-tube protocol with built-in molecular barcoding to enable accurate estimation of transcript abundance. To achieve that, we synthesized oligonucleotide-modified dideoxynucleotides which enable the generation of cDNA libraries at the reverse transcription step. We validated the performance of MTAS-seq on well-characterized reference bulk RNA and further tested it with eukaryotic cell lysates. |
| Published |
Philadelphia : Taylor & Francis Inc |
| Type |
Journal article |
| Language |
English |
| Publication date |
2022 |
| CC license |
|
