Journal: PLoS ONE

Article Title: The Characterization of Novel Tissue Microbiota Using an Optimized 16S Metagenomic Sequencing Pipeline

doi: 10.1371/journal.pone.0142334

Figure Lengend Snippet: Replicability and reproducibility. (a) Stacked bar charts showing the actual relative abundance of bacterial families in the staggered and even BEI genomic mock communities and the measured relative abundance in triplicates of the families obtained with the MiSeq sequencing pipeline. (b) Stacked bar charts showing the relative abundance of bacterial families obtained by sequencing of triplicates of fecal samples and ileum mucosa samples collected from three different mice (Sample 1, 2 and 3). (c) Stacked bar charts showing the relative abundance of bacterial families obtained by sequencing of three samples of mouse ileum mucosa in six to seven runs each with different parameters described in the legend at the bottom left: different runs, new libraries from same extracted DNA or the same libraries already prepared, different MiSeq kit generation and different sequencers. Two different experimenters performed the different runs and reagent batch numbers (including Taq polymerase) varied from run to run.

Article Snippet: This PCR was performed using 2 U of a DNA-free Taq DNA Polymerase and 1x Taq DNA polymerase buffer (MTP Taq DNA Polymerase, Sigma).

Techniques: Sequencing, Mouse Assay