Use of ATCC[R] MSA-2002[TM] for Validation of Extraction and Amplification Techniques in 16S Microbial Community Profiling

The implementation of microbiome analysis is an emerging area of focus in forensic research. However, microbiome analysis is not well validated for use in forensic analyses and there is no standard protocol in place. In this research a comprehensive analysis of extraction and amplification techniques employed during investigation of microbial communities was performed using ATCC[R] MSA-2002[TM] as a mock microbial community. Comparison of DNA extraction protocols was performed followed by an analysis of commercially-available polymerases. Samples were pooled for sequencing of the V4 region of 16S ribosomal RNA gene using the Illumina MiSeq System, and subsequently analyzed for community composition. The results were compared with the known genomic data of the mock microbial community and statistical methods were employed to determine the extent of deviation.