organism: coral metagenome
attributes.tag: investigation_type
attributes.value: miens-survey
attributes.tag: project_name
attributes.value: Cold-Water Coral Microbiomes (Lophelia pertusa) from Atlantic Ocean
attributes.tag: collection_date
attributes.value: 2018-08-29
attributes.tag: env_package
attributes.value: host-associated
attributes.tag: host_common_name
attributes.value: Desmophyllum pertusum
attributes.tag: host_taxid
attributes.value: 174260
attributes.tag: env_biome
attributes.value: ocean
attributes.tag: env_feature
attributes.value: cold-water coral Lophelia pertusa
attributes.tag: env_material
attributes.value: coral tissue
attributes.tag: geo_loc_name
attributes.value: Atlantic Ocean: Cape Fear Slide
attributes.tag: lat_lon
attributes.value: 33.572561 N 76.465048 W
attributes.tag: depth
attributes.value: 381 m
attributes.tag: temp
attributes.value: 6.99 deg C
attributes.tag: salinity
attributes.value: 35.54
attributes.tag: samp_collect_device
attributes.value: Manned submersible
attributes.tag: samp_mat_process
attributes.value: Preserved using RNAlater overnight at 4 ûC before extended storage in -20 ûC
attributes.tag: nucl_acid_ext
attributes.value: Extracted DNA from 0.15 g of coral tissue following manufacturer's protocol for MO BIO PowerBiofilm DNA Isolation Kit with the following modifications. Replaced Step 1 with a 2-mL rinse with cold sterile 1X PBS, then centrifuged at 6000xg in a refrigerated microcentrifuge for 15 min and removed PBS. Step 5 was accomplished by homogenization for 30 sec at 4200 rpm using a Mini-BeadBeater.
attributes.tag: target_gene
attributes.value: 16S rRNA
attributes.tag: target_subfragment
attributes.value: V4
attributes.tag: nucl_acid_amp
attributes.value: To target the V4 region of the 16S rRNA gene, a fusion primer set was constructed using primers 515F and 806RB (Apprill et al. (2015), doi: 10.3354/ame01753) along with adapters, indices, linkers, and pads in accordance with the dual-index sequencing strategy of Kozich et al. (2013, doi: 10.1128/aem.01043-13). Amplicons were purified, quantified, and pooled in equal concentrations.
attributes.tag: pcr_primers
attributes.value: FWD (515F):5_ GTGCCAGCMGCCGCGGTAA, REV (806RB):5_ GGACTACNVGGGTWTCTAAT
attributes.tag: seq_meth
attributes.value: Illumina MiSeq with v2 chemistry generating paired-end 250-bp reads
attributes.tag: BioSampleModel
attributes.value: MIMARKS.survey
attributes.tag: BioSampleModel
attributes.value: MIGS/MIMS/MIMARKS.host-associated