Amplicon-Seq of pmoA: Elba methane SIP T0 sediment replicate B DNA
The pmoA gene encoding the -subunit of particulate methane monooxygenase was amplified by nested PCR using primer pairs A189F/A682R and A189F/mb661R as described in Horz et al. 2005, AEM, 71(5):2642-52. Triplicate PCR products were combined and purified using 10 g of linear polyacrylamide and 270 l of polyethylene glycol-NaCl 6000 solution. Purified amplicons were subjected to 454 pyrosequencing (GS FLX Titanium system, MR DNA, Shallowater, TX, USA).