title: Bacterial 16S amplicon sequencing for mealworm feces
design: he feces collected from T. molitor larvae fed bran (control group, CG) or Styrofoam (treatment group, TG) were sent to Guandong Magigene Technology Co., Ltd. and Sangon Biotechnology (Shanghai) Co., Ltd., respectively. The near full-length 16S rRNA gene (V1-V9) amplicon sequencing were using PacBio Sequel IIe sequencer with the primer set of 27F (5-AGRGTTYGATYMTGGCTCAG-3) and 1492R (5-RGYTACCTTGTTACGACTT-3). The pooled 16S amplicons (600 ng) were used as template to prepare SMRTbell library via SMRTbell Express template prep kit 2.0 (PacBio, Menlo Park, CA, USA), according to the manufacturers instructions. After steps for damage repairing and end repairing, the inserts were ligated to adapters. The SMRT sequencing was performed on SMRT 8M Cell (PacBio, Menlo Park, CA, USA) with chemistry version 2 on PacBio Sequel IIe sequencer. A primary filtering analysis was achieved on the Sequel IIe System, and the secondary analysis was completed using the SMRT analysis pipeline version 11.0.0.