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Bio Protoc. 2019 Apr 20;9(8). pii: e3219. doi: 10.21769/BioProtoc.3219.

Northern Blot with IR Fluorescent Probes: Strategies for Probe Preparation.

Fields C1,2, Sheng P1,2, Miller B1, Wei T1, Xie M1,2,3.

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Department of Biochemistry and Molecular Biology, University of Florida, Gainesville, United States.
UF Health Cancer Center, University of Florida, Gainesville, United States.
UF Genetics Institute, University of Florida, Gainesville, United States.


Northern blot is a molecular biology technique that can detect, quantify, and determine the molecular weight of RNA. Recently, we published a protocol utilizing near-infrared (IR) fluorescent probes in Northern blot (irNorthern). Our method is as sensitive as other non-radioactive methods but is more straightforward and versatile. Additionally, we found that IR-labeled probes can be used to multiplex or detect different species of RNA at the same time. Here we describe three methods for generating an IR-labeled probe as well as how to perform irNorthern blot. In conclusion, our irNorthern protocol offers a convenient method for RNA detection.


Near infrared; Non-radioactive; Northern blot; RNA detection; irNorthern

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