title: Amplicon Seq of shrimp gut PMC
design: I extracted microbial DNA from shrimp gut using the TIANamp Bacteria DNA Kit (TIANGEN, China) immediately after harvesting. I amplified the 16S rRNA gene targeting the V3V4 region with primers recommended by Klindworth et al. (2013). The primers included Illumina adapter overhangs for downstream library preparation. The forward primer was 5-TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGCCTACGGGNGGCWGCAG-3, and the reverse primer was 5-GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAGGACTACHVGGGTATCTAATCC-3. I selected an insert size of ~550 bp to ensure at least 50 bp overlap for paired-end 2300 bp sequencing. After PCR amplification and adapter incorporation, I prepared the libraries for Illumina sequencing to generate high-quality paired-end reads for metagenomic analysis.