16S rRNA gene from Guaymas Basin propane oxidizing enrichment
Approximately 15mL of the enrichment culture was collected and centrifuged for 30 minutes at 5000g and 10C. The supernatant was removed and the resulting cell pellet was extracted using the manufacturers Gram positive protocol with the Qiagen DNEasy Blood and Tissue Kit (Cat. #69504, Qiagen, Hilden, Germany). 16S rRNA was amplified in duplicate using the 515F and 926R primers and the Q5 Hot Start Master Mix (Cat. M0492S, New England Biolabs). Libraries were barcoded with the Illumina Nextera XT index2 primers and amplified using the Q5 Hot Start PCR mixture. The sample was sequenced by Laragen (Culver City, CA, USA)