✅ High Sensitivity: Click chemistry reaction efficiency is extremely high, with clear signals and low background noise.

✅ High Specificity: Ethynyl and azide groups are almost absent in biological systems, resulting in extremely low reaction background.

✅ Good Biocompatibility: Reaction conditions are mild, causing minimal damage to RNA.

✅ Versatility: By replacing different azide reporter molecules, various purposes such as detection (fluorescence) and enrichment (biotin) can be achieved.

✅ Safety: No need to use radioactive labels, making operations simpler and safer.

One click, instantly illuminates the RNA synthesis process.

Principle: 5-Ethynyl-UTP can be directly incorporated by RNA polymerase into nascent RNA chains, replacing part of the natural UTP.

Detection Method: After incorporation, using its terminal alkyne group (–C≡CH), it can react with azide-labeled fluorescent dyes or biotin via click chemistry, enabling RNA visualization or enrichment.

Typical Reaction System: Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC).

Detection Forms:

• In vitro transcription system: 5-Ethynyl-UTP can directly replace a portion of UTP (typically 5–20% replacement ratio).

• Intracellular labeling: Commonly used 5-Ethynyl-uridine (EU) enters cells and is phosphorylated in vivo to 5-Ethynyl-UTP.

• Detection chemical reagents: Recommended to pair with Alexa Fluor 488-azide (fluorescence) or biotin-azide (enrichment).