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Microbial (16S) diversity in sediments of the former subglacial Hodgson lake (Antarctica)

Последняя версия опубликована SCAR - Microbial Antarctic Resource System 19 марта 2019 г. SCAR - Microbial Antarctic Resource System
Дата публикации:
19 марта 2019 г.
Лицензия:
CC-BY 4.0

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Описание

Amplicon sequencing dataset (454 pyrosequencing) of microorganisms (16S ssu rRNA gene) in sediments of the former subglacial Hodgson lake (Antarctica). Exploratory study (1 sample).

Версии

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Как оформить ссылку

Исследователи должны дать ссылку на эту работу следующим образом:

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

Права

Исследователи должны соблюдать следующие права:

Публикующей организацией и владельцем прав на данную работу является SCAR - Microbial Antarctic Resource System. This work is licensed under a Creative Commons Attribution (CC-BY) 4.0 License.

Регистрация в GBIF

Этот ресурс был зарегистрирован в GBIF, ему был присвоен следующий UUID: b315baf6-9fed-405d-a3ee-d585aae81a1c.  SCAR - Microbial Antarctic Resource System отвечает за публикацию этого ресурса, и зарегистрирован в GBIF как издатель данных при оподдержке Scientific Committee on Antarctic Research.

Ключевые слова

Metadata

Контакты

Кто является создателем ресурса:
-

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

Кто может ответить на вопросы о ресурсе:

David Pearce
British Antarctic Survey
Cambridge
GB

Кем заполнены метаданные:
-

Maxime Sweetlove
Research assistent
Royal Belgian Institute of natural sciences
Rue Vautier 29
1000 Brussels
BE

Кто еще связан с данным ресурсом:

User

Географический охват

Lake Hodgson, Antarctica

Ограничивающие координаты Юг Запад [-72,009, -68,462], Север Восток [-72,009, -68,462]

Таксономический охват

Bacteria and Archaea (16S ssu rRNA gene)

Domain Bacteria (Bacteria), Archaea (Archaea)

Временной охват

Дата начала 2000-01-01

Данные проекта

Описание отсутсвует

Название Hodgson Lake metagenome
Финансирование Funding was provided by theNatural Environment Research Council funding through the British Antarctic Survey and the Antarctic Funding Initiative.

Исполнители проекта:

David Pearce

Методы сбора

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.

Охват исследования Lake Hodgson, Antarctica

Описание этапа методики:

  1. 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.
  2. 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).

Библиографические ссылки

  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.

Дополнительные метаданные

Альтернативные идентификаторы b315baf6-9fed-405d-a3ee-d585aae81a1c
https://ipt.biodiversity.aq/resource?r=hodgson_lake_microbes