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DNA purity and integrity were analyzed using agarose gel electrophoresis. The DNA concentration was precisely quantified using a Qubit instrument. Qualified DNA samples were randomly fragmented into approximately 350 bp fragments using a Covaris ultrasonicator. Subsequently, the fragmented DNA underwent end repair, A-tailing, sequencing adapter ligation, purification, and PCR amplification to complete the library preparation. Following library construction, initial quantification was performed using a Qubit 2.0, and the library was diluted to a concentration of 2 ng/μl. The insert size of the library was then determined using an Agilent 2100, and once it met the expected size range, the library’s effective concentration was accurately quantified using Q-PCR (library effective concentration > 3 nM). After passing library quality control, different libraries were pooled based on their effective concentrations and the desired data output for Illumina PE150 sequencing. |
OTHER |
METAGENOMIC |
PCR |
PAIRED
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