Vibrio in Yucatan Coastal Marshes-CECS
"Bacterial tag-encoded FLX amplicon pyrosequencing (bTEFAP) will be performed as described previously using Gray28F 5?TTTGATCNTGGCTCAG and Gray519r 5? GTNTTACNGCGGCKGCTG  (Dowd, Callaway et al. 2008; Dowd, Sun et al. 2008; Dowd, Sun et al. 2008; Dowd, Wolcott et al. 2008; Callaway, Dowd et al. 2009; Guerrero, Dowd et al. 2009; Guerrero, Dowd et al. 2009; Leake, Dowd et al. 2009; Li, Dowd et al. 2009; Sen, Ishak et al. 2009; Suchodolski, Dowd et al. 2009; Wolcott, Gontcharova et al. 2009; Wolcott, Gontcharova et al. 2009; Bailey, Dowd et al. 2010; Bailey, Walton et al. 2010; Callaway, Dowd et al. 2010; Finegold, Dowd et al. 2010; Gontcharova, Youn et al. 2010; Olafson, Lohmeyer et al. 2010; Pitta, Pinchak et al. 2010; Smith, Snow et al. 2010; Stephenson, Mfuna et al. 2010; Williams, Tedeschi et al. 2010; Andreotti, Perez de Leon et al. 2011; Handl, Dowd et al. 2011; Ishak, Plowes et al. 2011).  Initial generation of the sequencing library will utilize a one-step PCR with a total of 30 cycles, a mixture of Hot Start and HotStar high fidelity taq polymerases, and amplicons originating and extending from the 28F for bacterial diversity.  Tag-encoded FLX amplicon pyrosequencing analyses utilized Roche 454 FLX instrument with Titanium reagents, titanium procedures performed at the Research and Testing Laboratory (Lubbock, TX) based upon RTL protocols (www.researchandtesting.com)."