Microbes (Eukaryotes and Archaea) in sea water from Fildes Peninsula (King George Island, Antarctica)

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如何引用

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Luo W, Li H, Gao S, Yu Y, Lin L, Zeng T (2019): Microbes (Eukaryotes and Archaea) in sea water from Fildes Peninsula (King George Island, Antarctica). v1.2. SCAR - Microbial Antarctic Resource System. Dataset/Metadata. https://ipt.biodiversity.aq/resource?r=microbes_fildes_peninsula_antarctica&v=1.2

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此資料的發布者及權利單位為 SCAR - Microbial Antarctic Resource System。 This work is licensed under a Creative Commons Attribution (CC-BY 4.0) License.

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此資源已向GBIF註冊，並指定以下之GBIF UUID: a4af5ceb-4035-49f5-b41a-bb548307b4f8。  SCAR - Microbial Antarctic Resource System 發佈此資源，並經由Scientific Committee on Antarctic Research同意向GBIF註冊成為資料發佈者。

關鍵字

Metadata

聯絡資訊

地理涵蓋範圍

Fildes Peninsula, King George Island, Antarctica

分類群涵蓋範圍

microbial Eukaryotes were sampled based on marker gene amplification (18S ssu rRNA gene)

Archaea were sampled based on marker gene amplification

時間涵蓋範圍

計畫資料

無相關描述

參與計畫的人員:

取樣方法

1 L of surface sea water from each station was collected and prefiltered through a 20-µm mesh sieve to remove most of the mesozooplankton and large particles, and then directly filtered through a 0.2-µm pore size nucleopore membrane filter (Whatman). The filters were frozen at −80 °C in cetyltrimethylammonium bromide (CTAB) buffer until laboratory experiments were carried out.

方法步驟描述:

1. DNA extraction was performed as described by Luo et al. (2009).
2. Polymerase chain reaction (PCR) was performed using primers with barcodes flanking the hypervariable V4 region of the 18S rRNA gene: 3NDf with the reverse primer V4_euk_R2. PCR was conducted in 20 μL reactions with 0.2 μM each of the primers, ~10 ng of template DNA, 1 × PCR buffer, and 2.5 U of Pfu DNA Polymerase (Promega, USA). The amplification programme consisted of an initial denaturation step at 95 °C for 2 min, followed by 30 cycles at 95 °C for 30 s, 55 °C for 30 s, and 72 °C for 30 s, and a final extension of 72 °C for 5 min. PCR products were pooled and purified using a DNA gel extraction kit (Axygen, Hangzhou, China). The DNA concentration of each PCR product was determined using a Quant-iT PicoGreen double-stranded DNA assay (Invitrogen, Germany) and was quality controlled on a TBS-380 Mini-Fluorometer (Turner Biosystems, Sunnyvale, CA, USA). Finally, amplicons of all samples were pooled in equimolar concentrations.
3. 18S rRNA amplification and sequencing on the Illumina MiSeq2000 were done by following the standard protocols of Earth Microbiome Project (EMP) (Caporaso et al. 2012).

引用文獻

1. Luo, W., Li, H., Gao, S., Yu, Y., Lin, L., & Zeng, Y. (2016). Molecular diversity of microbial eukaryotes in sea water from Fildes Peninsula, King George Island, Antarctica. Polar Biology, 39(4), 605-616. https://doi.org/10.1007/s00300-015-1815-8

額外的詮釋資料