16S rRNA sequencing of extracellular vesicles in patients with acute cholecystitis
After combining 40-50 amplicons into sub-libraries, each sub-library underwent purification utilizing the Qiaquick PCR purification kit (QIAGEN, Germany) as per the manufacturer's guidelines. Subsequently, multiple sub-libraries were pooled to create the final library. Subsequently, further purification from primer dimers was conducted using Agencourt AMPure XP (Beckman Coulter) at a volumetric ratio of 1:0.85 (Library: beads), followed by supplementation of the library with 15% PhiX (8 pM).