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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.

Author information

1
Department of Biochemistry and Molecular Biology, University of Florida, Gainesville, United States.
2
UF Health Cancer Center, University of Florida, Gainesville, United States.
3
UF Genetics Institute, University of Florida, Gainesville, United States.

Abstract

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.

KEYWORDS:

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

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