High throughput method of 16S rRNA gene sequencing library preparation for plant root microbial community profiling.

ABSTRACT: Microbiota are a major component of agroecosystems. Root microbiota, which inhabit the inside and surface of plant roots, play a significant role in plant growth and health. As next-generation sequencing technology allows the capture of microbial profiles without culturing the microbes, profiling of plant microbiota has become a staple tool in plant science and agriculture. Here, we have increased sample handling efficiency in a two-step PCR amplification protocol for 16S rRNA gene sequencing of plant root microbiota, improving DNA extraction using AMPure XP magnetic beads and PCR purification using exonuclease. These modifications reduce sample handling and capture microbial diversity comparable to that obtained by the manual method. We found a buffer with AMPure XP magnetic beads enabled

SUBMITTER: Kumaishi K 

PROVIDER: S-EPMC9652414 | biostudies-literature | 2022 Nov

REPOSITORIES: biostudies-literature