ArticlePlant methods2012
A cost-effective method for Illumina small RNA-Seq library preparation using T4 RNA ligase 1 adenylated adapters.
Article in Plant methods, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 39 papers.
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Who cites it
39 citing papers in PubMed.
- Evolutionarily New Solanaceous DCL2 Family Members Acquire Functions in Tomato.Molecular plant pathology · 2025Article
- Rapid discovery of functional RNA domains.Nucleic acids research · 2025Article
- Article
- Formamide significantly enhances the efficiency of chemical adenylation of RNA sequencing ligation adaptors.RNA (New York, N.Y.) · 2023Article
- Direct adenylation from 5'-OH-terminated oligonucleotides by a fusion enzyme containing Pfu RNA ligase and T4 polynucleotide kinase.Nucleic acids research · 2022Article
- Characterizing serotonin biosynthesis in Setaria viridis leaves and its effect on aphids.Plant molecular biology · 2022Article
- High-throughput sequencing application in the detection and discovery of viruses associated with the regulated citrus leprosis disease complex.Frontiers in plant science · 2022Article
- Article
- Transcriptome analysis provides insights into the responses of sweet potato to sweet potato virus disease (SPVD).Virus research · 2021Article
- A Small RNA-Seq Protocol with Less Bias and Improved Capture of 2'-O-Methyl RNAs.Methods in molecular biology (Clifton, N.J.) · 2021Article
- Applications of phage-derived RNA-based technologies in synthetic biology.Synthetic and systems biotechnology · 2020Review
- Deep Sequencing of Small RNAs in the WhiteflyInsects · 2020Article
- Revealing biomass heterosis in the allodiploid xBrassicoraphanus, a hybrid between Brassica rapa and Raphanus sativus, through integrated transcriptome and metabolites analysis.BMC plant biology · 2020Article
- Characterization of distinct strains of an aphid-transmitted ilarvirus (Fam. Bromoviridae) infecting different hosts from South America.Virus research · 2020Article
- Potato virus Y; the Andean connection.Virus evolution · 2019Article
- Article
- Complete Genome Sequence ofGenome announcements · 2018Article
- Systematic comparison of small RNA library preparation protocols for next-generation sequencing.BMC genomics · 2018Article
- Comprehensive transcriptome analyses reveal tomato plant responses to tobacco rattle virus-based gene silencing vectors.Scientific reports · 2017Article
- Capture, amplification, and global profiling of microRNAs from low quantities of whole cell lysate.The Analyst · 2017Article
Corrections and comments
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundDeep sequencing is a powerful tool for novel small RNA discovery. Illumina small RNA sequencing library preparation requires a pre-adenylated 3' end adapter containing a 5',5'-adenyl pyrophosphoryl moiety. In the absence of ATP, this adapter can be ligated to the 3' hydroxyl group of small RNA, while RNA self-ligation and concatenation are repressed. Pre-adenylated adapters are one of the most essential and costly components required for library preparation, and few are commercially available.
resultsWe demonstrate that DNA oligo with 5' phosphate and 3' amine groups can be enzymatically adenylated by T4 RNA ligase 1 to generate customized pre-adenylated adapters. We have constructed and sequenced a small RNA library for tomato (Solanum lycopersicum) using the T4 RNA ligase 1 adenylated adapter.
conclusionWe provide an efficient and low-cost method for small RNA sequencing library preparation, which takes two days to complete and costs around $20 per library. This protocol has been tested in several plant species for small RNA sequencing including sweet potato, pepper, watermelon, and cowpea, and could be readily applied to any RNA samples.
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Registered trials
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