ArticleCurrent issues in molecular biology2021
A Comprehensive Analysis of Northern versus Liquid Hybridization Assays for mRNAs, Small RNAs, and miRNAs Using a Non-Radiolabeled Approach.
Article in Current issues in molecular biology, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 24 citations in OpenAlex.
- RNA Labeling for Measuring RNA Endonucleolytic Cleavage.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Molecular arms races at the virus-host splicing interface and their pathogenic implications.Frontiers in microbiology · 2026Review
- Shaping Cell Identity: Global Transcriptome and Pathway Shifts during Mouse Mammary Epithelial Cell Differentiation.Computational and structural biotechnology journal · 2026Article
- Current updates regarding biogenesis, functions and dysregulation of microRNAs in cancer: Innovative approaches for detection using CRISPR/Cas13‑based platforms (Review).International journal of molecular medicine · 2025Review
- Quantitative and Multiplexing Analysis of MicroRNAs by Direct Full-Length Sequencing in Nanopores.Journal of the American Chemical Society · 2025Article
- Whole transcriptome sequencing reveals the functional regulation of chicken reproduction by lncRNAs, miRNAs, and mRNAs across the hypothalamus-pituitary-ovary axis.Frontiers in veterinary science · 2025Article
- Liquid and Solid Hybridization Methods to Detect RNAs.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Global Down-regulation of Gene Expression Induced by Mouse Mammary Tumor Virus (MMTV) in Normal Mammary Epithelial Cells.Viruses · 2023Article
- A New Biotin Labeling and High-Molecular-Weight RNA Northern Method and Its Application in Viral RNA Detection.Viruses · 2022Article
- Accurate detection of lung cancer-related microRNA through CRISPR/Cas9-assisted garland rolling circle amplification.Journal of thoracic disease · 2022Article
- Modified Northern blot protocol for easy detection of mRNAs in total RNA using radiolabeled probes.BMC genomics · 2022Article
- Nanoribbon Biosensor in the Detection of miRNAs Associated with Colorectal Cancer.Micromachines · 2021Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
Northern blotting (NB), a gold standard for RNA detection, has lost its charm due to its hands-on nature, need for good quality RNA, and radioactivity. With the emergence of the field of microRNAs (miRNAs), the necessity for sensitive and quantitative NBs has again emerged. Here, we developed highly sensitive yet non-radiolabeled, fast, economical NB, and liquid hybridization (LH) assays without radioactivity or specialized reagents like locked nucleic acid (LNA)- or digoxigenin-labeled probes for mRNAs/small RNAs, especially miRNAs using biotinylated probes. An improvised means of hybridizing oligo probes along with efficient transfer, cross-linking, and signal enhancement techniques was employed. Important caveats of each assay were elaborated upon, especially issues related to probe biotinylation, use of exonuclease, and bioimagers not reported earlier. We demonstrate that, while the NBs were sensitive for mRNAs and small RNAs, our LH protocol could efficiently detect these and miRNAs using less than 10-100 times the total amount of RNA, a sensitivity comparable to radiolabeled probes. Compared to NBs, LH was a faster, more sensitive, and specific approach for mRNA/small RNA/miRNA detection. A comparison of present work with six seminal studies is presented along with detailed protocols for easy reproducibility. Overall, our study provides effective platforms to study large and small RNAs in a sensitive, efficient, and cost-effective manner.
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Registered trials
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