16S rRNA Amplicon Sequencing of Liver Tissue in Diseased Procypris rabaudi
Sequencing libraries were constructed using the Illumina TruSeq Nano DNA LT Library Prep Kit according to the manufacturer's protocol. Briefly, the amplified products were subjected to end repair using the End Repair Mix 2 to remove 5' overhangs and add phosphate groups, followed by 3' end filling. An 'A' base was then added to the 3' ends to prevent fragment self-ligation and enable ligation to T-overhang adapters. Library-specific index adapters were ligated to the 5' ends to facilitate flow cell immobilization. The ligation products were purified using BECKMAN AMPure XP Beads to remove adapter dimers. Adapter-ligated fragments were amplified by PCR to enrich the library templates, followed by a second round of purification with AMPure XP Beads. Final size selection and purification were performed by 2% agarose gel electrophoresis.