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. 2025 Oct 27;17:2441–2457. doi: 10.2147/CMAR.S540752

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© 2025 Lv et al.

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Establishment and evaluation of methods to detect tRF5-23-GlyTCC-2. (A and B) Stability assessment: Room temperature and repeated freeze-thaw experiments showed no significant changes in the expression levels of tRF5-23-GlyTCC-2. (ns, p>0.05) (C) Agarose gel electrophoresis confirmed the RT-qPCR product of tRF5-23-GlyTCC-2 as a single band of approximately 80 bp. The red box represents the sequence of tRF5-23-GlyTCC-2. (D) Sanger sequencing of the RT-qPCR product verified the presence of the full-length sequence of tRF5-23-GlyTCC-2. (E) Amplification curve analysis: Red lines indicate tRF5-23-GlyTCC-2, and blue lines represent RNU6B. (F) Melting curve analysis: Left lines represent RNU6B, and right lines correspond to tRF5-23-GlyTCC-2.