5’-transfer RNA halve-lysine-CTT as a promising biomarker for early detection of hepatocellular carcinoma

Figure 2 Validation of 5’-transfer RNA halve-lysine-CTT as a biomarker. A: Schematic representation of the 5’-transfer RNA halve (tiRNA)-lysine (Lys)-CTT cleavage site, located above the anticodon loop; B and C: Stability of 5’-tiRNA-Lys-CTT assessed by quantitative polymerase chain reaction (qPCR) after exposure to varying durations of incubation at ambient temperature and multiple freeze-thaw cycles demonstrating consistent expression; D: Agarose gel electrophoresis of qPCR products showing a single band around 80 base pairs; E: Sanger sequencing confirmation of the qPCR product, demonstrating the full-length sequence of 5’-tiRNA-Lys-CTT; F: Representative melting curves of 5’-tiRNA-Lys-CTT, exhibiting a single peak, indicating high specificity. tiRNA: Transfer RNA halve; Lys: Lysine; CT: Cycle threshold.