KaOct12-bacteria
Bacterial and archaeal Tag-encoded FLX amplicon pyrosequencing (bTEFAP and aTEFAP, respectively), analyses by means of a Roche 454 FLX instrument with titanium reagents, were performed at the Research and Testing Laboratory (Lubbock, TX, USA) as described previously (Dowd et al., 2009; Callaway et al., 2010). The PCR primers for FLX amplicon pyrosequencing were chosen to span the variable V1–V3 regions in the 16S rRNA gene: 27F (5′-GAGTTTGATCNTGGCTCAG-3′) and 519R (5′-GWNTTACNGCGGCKGCTG-3′) for bacteria and ARCH 349F (5′-GYGCASCAGKCGMGAAW-3′) and ARCH 806R (5′-GGACTACVSGGGTATCTAAT-3′) for archaea.