miRNA-seq of homo sapiens: plasma exosome
Plasma samples were obtained from patients patients who underwent percutaneous coronary intervention (PCI) to treat CTOand AMI during coronary angiography. All patients of the CTO and AMI groups underwent exosome isolation from plasma samples and miRNA expression profiling. Total RNA was isolated from exosomes into the TRIzol LS reagent (Ambion, Life Technology, Carlsbad, CA, USA) according to the manufacturers instructions. RNA quality was assessed with the aid of an Agilent 2100 Bioanalyzer fitted with the RNA 6000 Pico Chip (Agilent Technologies, Amstelveen, the Netherlands); RNA quantification employed the Qubit assay system (Thermo Fisher Scientific, Waltham, MA, USA). Libraries of control and test RNAs were constructed using NEBNext Multiplex Small RNA Library Prep kits (New England BioLabs, Inc., Ipswich, MA, USA) according to the manufacturers instructions. The yields and size distributions of small RNA libraries were assessed with the aid of the Agilent 2100 Bioanalyzer instrument running the High-sensitivity DNA Assay (Agilent Technologies). High-throughput sequences were generated using the NextSeq 500 system employing single-end 75 sequencing.