Work

Northern Blot with IR Fluorescent Probes: Strategies for Probe Preparation

Year: 2019

Type: article

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.

Source: BIO-PROTOCOL

DOI: 10.21769/bioprotoc.3219

Authors: Christopher Fields, Peike Sheng, Bret Miller, Tianqi Wei, Mingyi Xie

Institutions: University of Florida, UF Health Cancer Center

Language: English

Topic: Molecular Biology Techniques and Applications

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: bronze

Funders: National Cancer Institute, National Institute of General Medical Sciences

Awards: Noncanonical microRNA biogenesis and function in a gamma herpesvirus and mammals, Molecular mechanisms for regulating microRNA levels in metazoans