Quantifying Bias in 16S rRNA Experiments due to DNA Extraction, PCR Amplification, and Sequencing and Classification. mixed culture metagenome

The goals of this project are to quantify and characterize bias in 16S rRNA Studies due to a particular choice of DNA extraction, PCR amplification, and sequencing and taxonomic classification protocols. Labs will make choices in protocol based on the environment of interest, but bias will always remain. This experiment is designed to quantify the differences between actual and observed community compositions when equal numbers of 1) cells, 2) DNA, and 3) PCR product are blended. By comparing the results of the experiments, one can quantify the contribution to bias of each step. Also, one can build models to predict true community composition based on the observed community composition.