STAR Protocols (Mar 2023)
Pseudomonas-specific 16S rRNA insect gut-microbiome profiling using next-generation sequencing
Abstract
Summary: We present a detailed protocol for Pseudomonas-specific 16S rRNA gut-microbiome profiling of brown planthopper (BPH) populations collected across changing climates and geographical locations using next-generation sequencing. We provide a technique for comparative analysis of Pseudomonas species structure and composition across BPH populations. Additionally, using qPCR we quantify the titers of Pseudomonas species in BPH. This protocol can be adopted for analyzing microbiome dynamics and monitoring populations of other pests, a crucial aspect for understanding their biodiversity, speciation, and adaptations.For complete details on the use and execution of this protocol, please refer to Gupta et al. (2022).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.