a26cdb5b-ab4b-48b7-b2a4-8d4c4b0e0671
Planktonic 18S rRNA gene sequences (V4 region) from water samples collected around an Acropora pulchra colony (Tema'e Beach reef, Mo'orea)
In this work, samples for DNA collection from waters around an Acropora pulchra colony were taken in April 25th, 2018 (12:00 local time) at the Tema'e beach reef of Mo'orea (French Polynesia). Water samples were withdrawn from three different local sites around the colony, at a depth of ~1 m: (IN) between the living branches, ~1 cm next to the polyps; (AL) deeper between the branches, ~1 cm next to the coral skeleton colonized by turf algae; and (OUT) 2 meters down current from the patch over a sandy bottom ~2 m deep. At that sampling point, 0.8 L (IN and AL) and 2.5 L (OUT) seawater were sampled and were prefiltered through 200-µm mesh. The microbial biomass was then collected on 0.2 µm pore-size, 47 mm diameter polycarbonate filters using a peristaltic pump. The filters were flash frozen in liquid N2 and stored at -80°C. Total DNA was extracted using the phenol-chloroform protocol as described in Massana et al. (1997). Eukaryotic diversity was determined by amplicon sequencing of the V4 regions of the 18S rDNA genes using the Illumina MiSeq platform and paired-end reads (2 x 250 bp). PCR amplifications were done using the eukaryotic universal primers V4F (5'-CCAGCASCYGCG GTAATTCC-3') and V4R (5'-ACTTTCGTTCTTGATYRR-3') (Balzano et al. 2015). All samples were sequenced at the Research and Testing Laboratories (RTL, Lubbock, TX, USA).  References:  Balzano, S., Abs, E., and Leterme, S. (2015). Protist diversity along a salinity gradient in a coastal lagoon. Aquat. Microb. Ecol. 74, 263–277. Massana, R., Murray, A. E., Preston, C. M., and DeLong, E. 1302 F. (1997). Vertical distribution and phylogenetic characterization of marine planktonic Archaea in the Santa Barbara Channel. Appl. Environ. Microbiol. 63, 50–56.
In this work, samples for DNA collection from waters around an Acropora pulchra colony were taken in April 25th, 2018 (12:00 local time) at the Tema'e beach reef of Mo'orea (French Polynesia). Water samples were withdrawn from three different local sites around the colony, at a depth of ~1 m: (IN) between the living branches, ~1 cm next to the polyps; (AL) deeper between the branches, ~1 cm next to the coral skeleton colonized by turf algae; and (OUT) 2 meters down current from the patch over a sandy bottom ~2 m deep. At that sampling point, 0.8 L (IN and AL) and 2.5 L (OUT) seawater were sampled and were prefiltered through 200-µm mesh. The microbial biomass was then collected on 0.2 µm pore-size, 47 mm diameter polycarbonate filters using a peristaltic pump. The filters were flash frozen in liquid N2 and stored at -80°C. Total DNA was extracted using the phenol-chloroform protocol as described in Massana et al. (1997). Eukaryotic diversity was determined by amplicon sequencing of the V4 regions of the 18S rDNA genes using the Illumina MiSeq platform and paired-end reads (2 x 250 bp). PCR amplifications were done using the eukaryotic universal primers V4F (5'-CCAGCASCYGCG GTAATTCC-3') and V4R (5'-ACTTTCGTTCTTGATYRR-3') (Balzano et al. 2015). All samples were sequenced at the Research and Testing Laboratories (RTL, Lubbock, TX, USA).  References:  Balzano, S., Abs, E., and Leterme, S. (2015). Protist diversity along a salinity gradient in a coastal lagoon. Aquat. Microb. Ecol. 74, 263–277. Massana, R., Murray, A. E., Preston, C. M., and DeLong, E. 1302 F. (1997). Vertical distribution and phylogenetic characterization of marine planktonic Archaea in the Santa Barbara Channel. Appl. Environ. Microbiol. 63, 50–56.