The linear staple RNA was mixed with splint RNA at a ratio of 1:1.5 and then added 1 × T4 RNA ligase buffer. The solution was incubated at 95 °C for 5 min, then slowly cooled down to room temperature. After low-speed centrifugation, T4 RNA ligase was added to the mixture to incubate at 37 °C for 4 h, followed by heating at 65 °C for 15 min to inactivate the ligase.
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