BLESS induced sputum
Bacterial tag-encoded FLX amplicon pyrosequencing (bTEFAP) was performed as described previously using Gray28F 5'-TTTGATCNTGGCTCAG-3' and Gray519r 5'-GTNTTACNGCGGCKGCTG-3'). Initial generation of the sequencing library involved a one-step PCR of 30 cycles, using a mixture of Hot Start and HotStar high fidelity Taq DNA polymerase, as described previously (4). 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) using RTL protocols (www.researchandtesting.com).