Use of pyrosequencing and DNA barcodes to monitor variations in Firmicutes and Bacteroidetescommunities in the gut microbiota of obese humans

Table 4 Determination of F/B ratios using pyrosequencing and clonal sequencing

Sample	Sequencing method	Estimation method	F	B	ratio F/B
Brain abscess pus	16S rRNA P	Barcode hits	39	41	0.95
	16S rRNA P	RDP-II classifier	521	509	1.02
	16S rRNA CS	Number of clone sequences	49	50	0.98
Cystic fibrosis sputum	16S rRNA P	Barcode hits	34	2	17
	16S rRNA CS	Number of clone sequences	16	1	16
Caecal content	MP	Barcode hits	79	33	2.39
		% of total 16S rRNA sequences	60.70	28.41	2.14

P, CS and MP mean Pyrosequencing, Clonal Sanger Sequencing and Metagenome Pyrosequencing, respectively. F and B mean Firmicutes phylum and Bacteroidetes phylum, respectively. The numbers of clone sequences assigned to the Firmicutes or Bacteroidetes phylum was performed using BLAST algorithm and Genebank database. For the metagenome pyrosequencing, the 16S rRNA fraction was found with BLAST algorithm and RDP-II database. RDP-II classifier was used as a control with a bootstrap confidence > 80%

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