Small RNA sequencing of human serum and sperm samples associated with adverse childhood experiences
Small RNA sequencing of human serum and sperm samples associated with adverse childhood experiences
Serum and sperm samples were collected from participants of the PLMS and CTQ cohorts according to the study protocol. Samples were anonymized before sequencing and downstream analysis. Total RNA was extracted from human serum and sperm samples using QIAzol (Qiagen) or TRIzol (Invitrogen) reagents according to the study protocol. Briefly, 1 ml of QIAzol or TRIzol was added to 200 µl of serum or to the sperm pellet, respectively. RNA extraction was performed by chloroform-based phase separation followed by isopropanol precipitation. RNA concentration was assessed using a NanoDrop spectrophotometer. RNA quality and integrity were evaluated using the Agilent 2100 Bioanalyzer with the RNA 6000 Nano Kit. miRNA libraries were prepared using the QIAseq miRNA Library Kit according to the manufacturer's instructions. Libraries were generated from 200 ng of total RNA. Briefly, 3′ and 5′ adapters were ligated to miRNAs, followed by reverse transcription, cDNA purification, and PCR amplification. Library quality was evaluated using the Agilent 2100 Bioanalyzer with a High Sensitivity DNA chip. The mean library size was 174 bp.