GSM8794760: Pre_treatment_5; Homo sapiens; miRNA-Seq
Total RNA was extracted and quality controlled using NanoDrop and Agilent 2100 Bioanalyzer to assess RNA integrity, requiring RIN value ≥8 and 28S:18S ratio ≥1.5:1. Small RNA library was constructed using 1-10μg total RNA, including 3' adapter ligation, 5' adapter ligation, reverse transcription, and PCR amplification. The PCR products were separated by 6% polyacrylamide gel electrophoresis, and target fragments of 145-160bp were excised and recovered. Library quality control and quantification were performed using Bioanalyzer and qPCR, and the library concentration was adjusted to 2nM. Finally, cluster generation and sequencing were performed on the HiSeq 2500 platform, with sequencing quality assessed by First Base Report. Small RNA libraries were constructed using the Illumina platform. Library construction was performed using 1-10μg of quality-controlled total RNA (RIN≥8), including 3' adapter ligation, 5' adapter ligation, reverse transcription, and PCR amplification. The PCR products were separated by gel electrophoresis, and 145-160bp target fragments were recovered. After quality control and quantification, the library concentration was adjusted to 2nM for sequencing on the HiSeq 2500 platform.