# Protocol for the development, assembly, and testing of a synthetic skin microbial community

## TL;DR

This paper provides a detailed protocol for creating and analyzing a synthetic skin microbial community to study human skin health and disease.

## Contribution

The paper introduces SkinCom, a reproducible synthetic microbial community of nine dominant human skin bacteria, along with a full experimental and computational workflow.

## Key findings

- A synthetic microbial community (SkinCom) was developed using nine dominant human skin bacteria strains.
- The protocol includes steps for DNA extraction, library preparation, and multiomic analysis of the synthetic community.
- The application of SkinCom was demonstrated using an epicutaneous murine model.

## Abstract

A reproducible study system is essential for understanding the role of microbes in human skin health and disease. We present a protocol for constructing a synthetic microbial community (SkinCom) of nine strains dominant to native human skin microbiome. We describe steps for computing growth metrics, constructing communities, and extracting DNA and library preparation for shotgun sequencing. We detail steps for data preprocessing and analysis of community samples. We illustrate SkinCom’s application with an epicutaneous murine model and downstream multiomic analysis.

For complete details on the use and execution of this protocol, please refer to Lekbua et al.1

•Automated construction of a reproducible synthetic microbial community for human skin•Steps for computing microbial growth metrics for nine dominant human-skin bacteria•Extracting DNA for metagenomics and metatranscriptomics•Computational pipeline steps for preprocessing and analyzing FASTQ sequencing files

Automated construction of a reproducible synthetic microbial community for human skin

Steps for computing microbial growth metrics for nine dominant human-skin bacteria

Extracting DNA for metagenomics and metatranscriptomics

Computational pipeline steps for preprocessing and analyzing FASTQ sequencing files

Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

A reproducible study system is essential for understanding the role of microbes in human skin health and disease. We present a protocol for constructing a synthetic microbial community (SkinCom) of nine strains dominant to native human skin microbiome. We describe steps for computing growth metrics, constructing communities, and extracting DNA and library preparation for shotgun sequencing. We detail steps for data preprocessing and analysis of community samples. We illustrate SkinCom’s application with an epicutaneous murine model and downstream multiomic analysis.