Spectra were recorded with a Thermo Scientific BioMate 6 split beam UV/visible spectrophotometer. The concentrations of bacteriopheophytin a, bacteriochlorophyll a and spirilloxanthin in the acetone/methanol extracts were determined from the absorbance values obtained at 747, 771 and 475 nm, respectively, using the spectral reconstruction method of van der Rest and Gingras [60]. The GW-572016 price detection and identification of various cytochrome

types was done as reported selleckchem previously [8]. Chemotaxonomical characterization Cellular fatty acid patterns were determined from cells grown to stationary phase in SYPHC liquid medium or on Marine Agar 2216. The preparation and extraction of fatty acid methyl esters from biomass and their subsequent separation and identification by gas chromatography was done as described elsewhere [61]. Respiratory lipoquinone and polar lipid analyses were carried out by the Identification Service and Dr. B.J. Tindall, DSMZ, Braunschweig, Germany, according to the protocols given by the DSMZ Identification Service [62]. Detection of specific genes using PCR For the isolation of genomic DNA from strain Ivo14T and

further reference strains the MasterPure™ Gram Positive DNA Purification Kit from Epicentre (Madison, USA) was used according to the instructions provided by the manufacturer. Extracted genomic DNA was quantified using a NanoDrop ND1000 spectrophotometer (Peqlab; Erlangen, Germany). PCR amplification of genomic Vorinostat datasheet DNA was carried out using the HotMasterMix 2.5x from 5 PRIME (Hamburg, Germany) according to the manufacturer’s protocol or the Taq DNA polymerase

from Qiagen (Hilden, Germany) in reaction buffer Phloretin containing 200 μM (each) deoxynucleotide triphosphates (dNTPs), 1 μM (each) oligonucleotide primers and ca. 10 – 25 ng of genomic DNA in a final volume of 20 μl. PCR products were purified using the HiYield Gel/PCR clean-up and Gel-Extraction Kit (SLG; Gauting, Germany) according to the manufacturer’s protocol and visualized by gel electrophoresis (1% agarose). Finally, PCR products were sequenced using a BigDye Terminator v3.1 Cycle Sequencing kit (Life Technologies; Darmstadt, Germany) and an ABI 3730xl DNA Analyzer (Applied Biosystems; Darmstadt, Germany). Amplification of pufLM genes For detection of pufL and pufM genes in extracted DNA a PCR amplification was performed with two sets of degenerated primers (see Table  4). Sequences of the primer set pufLF2/pufMR2 were optimized to match known sequences of BChl a-containing members of the OM60/NOR5 clade. The amplification comprises the following program: an initial step at 98°C for 3 min and then 35 cycles at 98°C for 15 s, 56°C for 25 s and 72°C for 1.5 min. At the end a postelongation at 72°C for 10 min was carried out.