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Figure 2 | BMC Microbiology

Figure 2

From: Sampling and pyrosequencing methods for characterizing bacterial communities in the human gut using 16S sequence tags

Figure 2

Comparison of the recovery of different bacterial taxa with use of different stool storage and DNA isolation methods. 473,169 sequence reads were used to characterize the 57 communities analyzed. All subjects tested for each method were pooled for comparison (summarized in Additional File 1). Methods are numbered at the top of the heat map. For the heat map scale, the number beside each colored tile indicates the lower bound for the indicated interval. Taxa are mostly indicated at the genus level; raee taxa are pooled. A) Comparison of DNA isolation using the Qiagen stool kit (methods 1 and 2) to lysis by bead-beating in hot phenol (method 9). Six subjects were compared. B) Comparison of the Qiagen stool kit samples (methods 1 and 2) to the MoBio Powersoil kit (method 3). Three subjects were compared. C) Comparison of methods for storage of stool specimens. DNA was prepared from fresh samples (method 8), samples stored frozen at -80 for several days (methods 1 and 2), or samples stored at 4°C for 24 hr (method 4) or 48 hr (method 5). Three subjects were compared. D) Comparison of stool storage in PSP (method 6) to storage methods 1, 2, 4 and 5. All 10 subjects were compared. For A) and D), the methods were compared using the Wilcoxon signed rank test to identify bacterial groups that significantly changed in proportion. (* indicates P < 0.05). Numbers of samples were too low in B) and C) for statistical testing.

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