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• W2891235038 abstract "Northern blot analysis detects RNA molecules immobilized on nylon membranes through hybridization with radioactive 32 P-labeled DNA or RNA oligonucleotide probes. Alternatively, nonradioactive northern blot relies on chemiluminescent reactions triggered by horseradish peroxidase (HRP) conjugated probes. The use of regulated radioactive material and the complexity of chemiluminescent reactions and detection have hampered the adoption of northern blot techniques by the wider biomedical research community. Here, we describe a sensitive and straightforward nonradioactive northern blot method, which utilizes near-infrared (IR) fluorescent dye-labeled probes (irNorthern). We found that irNorthern has a detection limit of ∼0.05 femtomoles (fmol), which is slightly less sensitive than 32 P-Northern. However, we found that the IR dye-labeled probe maintains the sensitivity after multiple usages as well as long-term storage. We also present alternative irNorthern methods using a biotinylated DNA probe, a DNA probe labeled by terminal transferase, or an RNA probe labeled during in vitro transcription. Furthermore, utilization of different IR dyes allows multiplex detection of different RNA species. Therefore, irNorthern represents a more convenient and versatile tool for RNA detection compared to traditional northern blot analysis." @default.
• W2891235038 created "2018-09-27" @default.
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• W2891235038 date "2018-09-10" @default.
• W2891235038 modified "2023-10-17" @default.
• W2891235038 title "Near-infrared fluorescent northern blot" @default.
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• W2891235038 doi "https://doi.org/10.1261/rna.068213.118" @default.
• W2891235038 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/6239192" @default.
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