摘要：The purpose of this review is to provide medical researchers,especially those without a bioinformatics background,with an easy-to-understand summary of the concepts and technologies used in microbiome ***,we define primary concepts such as microbiota,microbiome,and ***,we discuss study design schemes,the methods of sample size calculation,and the methods for improving the reliability of *** emphasize the importance of negative and positive controls in this ***,we discuss statistical analysis methods used in microbiome research,focusing on problems with multiple comparisons and ways to compare β-diversity between ***,we provide step-by-step pipelines for bioinformatics *** summary,the meticulous study design is a key step to obtaining meaningful results,and appropriate statistical methods are important for accurate interpretation of microbiome *** step-by-step pipelines provide researchers with insights into newly developed bioinformatics analysis methods.