16S rRNA Full-Length-Seq of BSFL Guava-fed Frass
A group of black soldier fly larvae were fed with guava-based substrate. The excreta (frass) samples of the larvae are used for DNA extraction and sequencing. The Full-length 16S rRNA gene was amplified from extracted microbial genomic DNA using KAPA HiFi HotStart Ready Mix PCR kit (KAPA Biosystems). Each PCR product was analyzed by a D5000 screentape assay run on TapeStation 4200 (Agilent Technologies, USA) to check for size distribution and quantification. The pooled PCR product was purified with AMPure PB beads (PacBio) and converted to barcoded Single-molecule real-time (SMRT) libraries using the SMRTbell Express Template Prep Kit 2.0 (PacBio). Purified with AMPure PB beads (PacBio), the libraries were quantified by the Qubit 1X dsDNA High Sensitivity Kit and size checked by D5000 ScreenTape Assay. Followed by sequencing on PacBio Sequel IIe platform, the raw data was used to generate Circular consensus sequence (CCS) with the CCS module in SMRT Link (PacBio). Primers for full-length 16S rRNA gene are Forward (F27): AGRGTTYGATYMTGGCTCAG and Reverse (R1492): RGYTACCTTGTTACGACTT