Bacterial diversity in closed cryoconite holes in Southern Victoria Land (Antarctica)

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Webster-Brown J, Hawes I, Jungblut A, Wood S, Christenson H (2019): Bacterial diversity in closed cryoconite holes in Southern Victoria Land (Antarctica). v1.2. SCAR - Microbial Antarctic Resource System. Dataset/Metadata. https://ipt.biodiversity.aq/resource?r=bacteria_in_closed_cryoconite_holes_antarctica&v=1.2

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L’éditeur et détenteur des droits de cette ressource est SCAR - Microbial Antarctic Resource System. Ce travail est sous licence Creative Commons Attribution (CC-BY) 4.0.

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Cette ressource a été enregistrée sur le portail GBIF, et possède l'UUID GBIF suivante : f03a9318-a568-433a-9c5d-db0b88b7e2a4.  SCAR - Microbial Antarctic Resource System publie cette ressource, et est enregistré dans le GBIF comme éditeur de données avec l'approbation du Scientific Committee on Antarctic Research.

Mots-clé

Metadata

Contacts

Couverture géographique

Diamind and Koettliz Glaciers (Southern Victoria Land, Antarctica)

Couverture taxonomique

Bacteria were profiled based on amplicon sequencing of the 16S ssu rRNA gene

Méthodes d'échantillonnage

Access through the ice lid was accomplished using a 50-mm diameter Kovacs ice drill, which was cleaned between each hole with antibacterial wipes. Water samples were collected directly into three 60-mL centrifuge tubes for determination of major ions and isotopes, using a manual vacuum apparatus. A sample (1 L) was collected by vacuum into an acid-washed PVC bottle. A subsample (100 mL) filtered (Whatman GF/F) into an acid-washed polyethylene bottle and frozen for later nutrient analysis.

Description des étapes de la méthode:

1. DNA was extracted from sediment using the MoBio Power Soil kit (USA). Biosystems, USA). A region of the 16S rRNA gene covering the V3 and 4 sections was ampli ed by PCR (iCycler; Biorad) using bacterial-speci c primers 515F and 806R (Caporaso et al. 2011). PCR reactions were performed in 50 μL volumes with the reaction mixture containing; 45 μL of Platinu PCR SuperMix High Fidelity (Life Technologies, USA), 10 μM of each primer, and 10–20 ng of template DNA. The reaction mix- ture was held at 94◦C for 2 min followed by 27 cycles of 94◦C for 30 s, 54◦C for 30 s, 68◦C for 45 s, with a nal extension step at 68◦C for 5 min. PCR products were puri ed (Agencour AMPur XP Kit; Beckman Coulter, USA), quanti ed (Qubit 20 Fluorometer, Life Technologies, USA), diluted to 1 ng μL−1 and submitted to New Zealand Genomics Limited (Auckland, New Zealand) for library preparation. Libraries were sequenced on a MiSeq Illumina platform (2 × 250 reads).

Citations bibliographiques

1. Webster-Brown, J. G., Hawes, I., Jungblut, A. D., Wood, S. A., & Christenson, H. K. (2015). The effects of entombment on water chemistry and bacterial assemblages in closed cryoconite holes on Antarctic glaciers. FEMS microbiology ecology, 91(12). https://doi.org/10.1093/femsec/fiv144

Métadonnées additionnelles