Pyrosequencing of prokaryote: a new black smoker vent field at 15буS of the Southern Mid-Atlantic Ridge
Amplification of the 16S rRNA gene V5-V8 region (approx.600bp) was performed using the universal prokaryotic primers Uni787F (5'-ATTAGATACCCNGGTAG-3') and Uni1391R (5'-ACGGGCGGTGWGTRC-3') as previously described (Jaeschke et al., 2012). A set of 8 nucleotide (nt) barcodes was designed and added to the 5' end of Uni787F for multiplexing of the samples in pyrosequencing runs. The barcodes for each of samples were: TVG.10C, AGAGACAT; TVG.10V, AGATCACT; TVG.11, AGCGCTCT; TVG.12, AGTCAGCT; TVG.13, AGTCAGTG; Sshrp.1, AGTGCTCG; Sshrp.2, ATACGTCG; Sshrp.3, ATACGTGT; Lshrp.1, ATAGACTG; Lshrp.2, ATCAGTGT; Lshrp.3, ATCATCAT; Mshrp.1  ATCGAGCT; Mshrp.2, ATCTATCT; Mshrp.3, ATGATGAT; CTD10.2600, ACGAGATG; CTD10.2700 , ACGATCGT; CTD10.2400, ACGTGACT; MiMa.05, ACACATAT.