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

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

Bio-protocol (2019-08-21)
Christopher Fields, Peike Sheng, Bret Miller, Tianqi Wei, Mingyi Xie
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Sodium acetate, BioXtra, ≥99.0%
Sigma-Aldrich
Trizma® base, Primary Standard and Buffer, ≥99.9% (titration), crystalline
Sigma-Aldrich
Sodium citrate dihydrate, ≥99%, FG
Sigma-Aldrich
Urea, powder, BioReagent, for molecular biology, suitable for cell culture
Sigma-Aldrich
Magnesium chloride hexahydrate, BioXtra, ≥99.0%