ArticleRNA biology2020
Systematic assessment of commercially available low-input miRNA library preparation kits.
Article in RNA biology, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed.
- Unique circulating miRNAome signatures captured with distinct molecular states in Lynch syndrome and sporadic colorectal cancer.Molecular oncology · 2026Article
- Wide-spectrum profiling of plasma cell-free RNA and the potential for health-monitoring.RNA biology · 2025Article
- Optimizing Small RNA Sequencing for Salivary Biomarker Identification: A Comparative Study of Library Preparation Protocols.International journal of molecular sciences · 2025Article
- Guidelines for mitochondrial RNA analysis.Molecular therapy. Nucleic acids · 2024Review
- Enhancing Clinical Utility: Utilization of International Standards and Guidelines for Metagenomic Sequencing in Infectious Disease Diagnosis.International journal of molecular sciences · 2024Review
- Serum microRNA Profiles and Pathways in Hepatitis B-Associated Hepatocellular Carcinoma: A South African Study.International journal of molecular sciences · 2024Review
- Respiratory virus infections and adenovirus characteristics during acute exacerbation of chronic obstructive pulmonary disease.Technology and health care : official journal of the European Society for Engineering and Medicine · 2024Article
- In search of the ratio of miRNA expression as robust biomarkers for constructing stable diagnostic models among multi-center data.Frontiers in genetics · 2024Article
- Reassessment of miRNA variant (isomiRs) composition by small RNA sequencing.Cell reports methods · 2023Article
- Comparative Analysis of Free-Circulating and Vesicle-Associated Plasma microRNAs of Healthy Controls and Early-Stage Lung Cancer Patients.Pharmaceutics · 2022Article
- APE1 controls DICER1 expression in NSCLC through miR-33a and miR-130b.Cellular and molecular life sciences : CMLS · 2022Article
- Development of Alzheimer's Disease Biomarkers: From CSF- to Blood-Based Biomarkers.Biomedicines · 2022Review
- Deciphering specific miRNAs in brain tumors: a 5-miRNA signature in glioblastoma.Molecular genetics and genomics : MGG · 2022Article
- Chemical capping improves template switching and enhances sequencing of small RNAs.Nucleic acids research · 2022Article
- A Unified Computational Framework for a Robust, Reliable, and Reproducible Identification of Novel miRNAs From the RNA Sequencing Data.Frontiers in bioinformatics · 2022Article
- Non-Coding RNAs in Preeclampsia-Molecular Mechanisms and Diagnostic Potential.International journal of molecular sciences · 2021Review
- Protocol to analyze circulating small non-coding RNAs by high-throughput RNA sequencing from human plasma samples.STAR protocols · 2021Article
- Quantitative mapping of the cellular small RNA landscape with AQRNA-seq.Nature biotechnology · 2021Article
- Single-cell microRNA sequencing method comparison and application to cell lines and circulating lung tumor cells.Nature communications · 2021Article
- Small RNA-Sequencing: Approaches and Considerations for miRNA Analysis.Diagnostics (Basel, Switzerland) · 2021Review
Corrections and comments
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Authors and funding
21 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
High-throughput sequencing is increasingly favoured to assay the presence and abundance of microRNAs (miRNAs) in biological samples, even from low RNA amounts, and a number of commercial vendors now offer kits that allow miRNA sequencing from sub-nanogram (ng) inputs. Although biases introduced during library preparation have been documented, the relative performance of current reagent kits has not been investigated in detail. Here, six commercial kits capable of handling <100ng total RNA input were used for library preparation, performed by kit manufactures, on synthetic miRNAs of known quantities and human total RNA samples. We compared the performance of miRNA detection sensitivity, reliability, titration response and the ability to detect differentially expressed miRNAs. In addition, we assessed the use of unique molecular identifiers (UMI) sequence tags in one kit. We observed differences in detection sensitivity and ability to identify differentially expressed miRNAs between the kits, but none were able to detect the full repertoire of synthetic miRNAs. The reliability within the replicates of all kits was good, while larger differences were observed between the kits, although none could accurately quantify the relative levels of the majority of miRNAs. UMI tags, at least within the input ranges tested, offered little advantage to improve data utility. In conclusion, biases in miRNA abundance are heavily influenced by the kit used for library preparation, suggesting that comparisons of datasets prepared by different procedures should be made with caution. This article is intended to assist researchers select the most appropriate kit for their experimental conditions.
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