GSM9090306: A885189, SALI; Homo sapiens; miRNA-Seq
Whole blood samples were collected by venipuncture into PAXGene Blood RNA tubes (BD, San Jose, CA, USA). Total RNA was extracted using on-column DNase digestion (Qiagen, Hilden, NRW, Germany). Excess globin transcripts were removed using the GLOBINclear kit (Ambion, Austin, TX, USA). RNA concentration was measured with a NanoDrop spectrophotometer, and RNA quality was evaluated with the Bioanalyzer 2100 (Agilent, Santa Clara, CA, USA) to determine its integrity, with confirmation by agarose gel electrophoresis. Samples were deemed suitable for downstream library preparation and sequencing only if they satisfied the following thresholds: concentration > 50 ng/μL, RNA integrity number (RIN) > 7.0, OD260/280 ratio > 1.8, and total RNA yield > 2 μg. Small RNA libraries were constructed using the TruSeq Small RNA Sample Prep Kits(Illumina, San Diego, USA) according to kit instructions. The specific procedures were as follows: 3 'splice sequence ligation, 5' splice sequence ligation, reverse transcription to generate cDNA strand, PCR amplification, electrophoresis purification of target fragment, sequencing. Instruments and Reagents for library quality Control, Inc Instrument: Bioanalyzer 2100 (Agilent, CA, USA) Library quality check Kit: High Sensitivity DNA Chip Kit (Agilent, CA, USA)