b315baf6-9fed-405d-a3ee-d585aae81a1c https://ipt.biodiversity.aq/resource?r=hodgson_lake_microbes Microbial (16S) diversity in sediments of the former subglacial Hodgson lake (Antarctica) David Pearce British Antarctic Survey
Cambridge GB
Dominic Hodgson British Antarctic Survey
Cambridge GB
Michael Thorne British Antarctic Survey
Cambridge GB
Gavin Burns British Antarctic Survey
Cambridge GB
Charles Cockell University of Edinburgh
Edinburgh GB
Maxime Sweetlove Royal Belgian Institute of natural sciences Research assistent
Rue Vautier 29 Brussels 1000 BE
msweetlove@naturalsciences.be
user 2019-03-19 eng Amplicon sequencing dataset (454 pyrosequencing) of microorganisms (16S ssu rRNA gene) in sediments of the former subglacial Hodgson lake (Antarctica). Exploratory study (1 sample). Metadata GBIF Dataset Type Vocabulary: http://rs.gbif.org/vocabulary/gbif/dataset_type.xml This work is licensed under a Creative Commons Attribution (CC-BY) 4.0 License. Lake Hodgson, Antarctica -68.462 -68.462 -72.009 -72.009 2000 2000 Bacteria and Archaea (16S ssu rRNA gene) domain Bacteria Bacteria domain Archaea Archaea unkown David Pearce British Antarctic Survey
Cambridge GB
Samples of frozen surface sediment (~0.5 g) were removed from sediment cores for DNA extraction. DNA extractions were carried out using the PowerSoil DNA isolation kit (Mo Bio, Carlsbad, CA, USA) following the manufacturers standard protocol. The resulting DNA was quantified using a Nanodrop spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA). Each extraction from 0.5 g sediment produced ~250 ng of DNA in 100 μL nuclease free water. 16S RNA gene amplification was carried out on sediment DNA obtained from the four depth sections A–C (15 ng input DNA) using forward primer 787f ATT AGA TAC CCN GGT AG and reverse primer 1492Rm GNT ACC TTG TTA CGA CTT at an annealing temperature of 50 °C to produce amplicons of 705 bp. PCR reactions were 15 μL and contained 1× PCR buffer, 2.0 mM MgCl2, 0.25 mM dNTPs, 0.25 mM each primer, 0.4 Units BioTaq (Bioline, London, UK) and ca. 15 ng template DNA. For Thermocycling: 1 ×95 °C for 50 s; 30 ×95 °C for 20 s, 50 °C for 30 s, 72 °C for 3 min (ramp 72 °C at 0.3 °C s−1); 1 ×72 °C for 10 min. Multiple PCR amplifications (24 in total) were cleaned and pooled to produce 900 ng of 16S enriched DNA for 454 library preparation. An amplicon library was generated using the Rapid Library Preparation kit and following the manufacturer’s recommendations in the GS FLX Titanium Series Rapid Library Preparation Method Manual (Roche). Briefly, the PCR amplicons were purified using AMPure beads (Agencourt Bioscience Corporation, Beverly, MA, USA), adaptors were blunt-end ligated to the fragment and the dsDNA amplicon library was quantitated via fluorometry using Quanti-iT Pico Green reagents (Invitrogen, Carlsbad, CA, USA). The library was then subjected to clonal amplification by emulsion PCR followed by pyrosequencing on a 454 GS FLX sequencer according to the manufacturer’s instructions (NEB NextQuick 454 library prep kit E6090). Lake Hodgson, Antarctica A 3.8 m sediment core was extracted at a depth of 93.4 m below the ice surface inside a virkon sterilized and polycarbonate lined core barrel. When taken all core samples remained within the core liners and were immediately frozen until sub-sampling in a clean class II microbiological safety cabinet in Cambridge. Gloves were used at all times and instruments used in laboratory manipulations were autoclaved. Hodgson Lake metagenome David Pearce Funding was provided by theNatural Environment Research Council funding through the British Antarctic Survey and the Antarctic Funding Initiative.
2019-02-12T11:38:59.927+01:00 dataset Pearce D, Hodgson D, Thorne M, Burns G, Cockell C (2019): Microbial (16S) diversity in sediments of the former subglacial Hodgson lake (Antarctica). v1.1. SCAR - Microbial Antarctic Resource System. Dataset/Metadata. https://ipt.biodiversity.aq/resource?r=hodgson_lake_microbes&v=1.1 Pearce, D. A., Hodgson, D. A., Thorne, M. A., Burns, G., & Cockell, C. S. (2013). Preliminary analysis of life within a former subglacial lake sediment in Antarctica. Diversity, 5(3), 680-702. b315baf6-9fed-405d-a3ee-d585aae81a1c/v1.1.xml