FR2A5
The sequencing library was prepared using the Illumina TruSeq Nano DNA LT Library Prep Kit. Amplified DNA fragments were first end-repaired using End Repair Mix 2 to remove 5 overhangs, add phosphate groups, and fill in 3 overhangs. An A-base was then added to the 3 ends to prevent self-ligation and ensure compatibility with sequencing adapters, which have a complementary T-base overhang at their 3 ends. Sequencing adapters containing library-specific index sequences were ligated to the 5 ends of the DNA fragments to enable attachment to the Flow Cell. Adapter-ligated fragments were purified using BECKMAN AMPure XP Beads to remove self-ligated adapters. The adapter-ligated DNA fragments were PCR-amplified to enrich the library templates, followed by further purification using BECKMAN AMPure XP Beads. Finally, 2% agarose gel electrophoresis was used for final fragment selection and purification.