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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
RESUMEN

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.

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Base Trizma®, Primary Standard and Buffer, ≥99.9% (titration), crystalline
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Urea, powder, BioReagent, for molecular biology, suitable for cell culture
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Citrato de sodio tribásico dihydrate, ≥99%, FG
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Cloruro de magnesio hexahydrate, BioXtra, ≥99.0%
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Sodium acetate, BioXtra, ≥99.0%