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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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Fildes Peninsula, King George Island, Antarctica
|Bounding Coordinates||South West [-62.2, -58.9], North East [-62.2, -58.9]|
microbial Eukaryotes were sampled based on marker gene amplification (18S ssu rRNA gene)
Archaea were sampled based on marker gene amplification
|Start Date / End Date||2013-01-17 / 2013-01-23|
No Description available
|Title||Microbes (Eukaryotes and Archaea) in sea water from Fildes Peninsula (King George Island, Antarctica)|
|Funding||This work was supported by the National High-Tech Research and Development Program of China (Grant Nos. 2012AA021706, 2013AA065805), National Natural Science Foundation of China (No. 41376191), Chinese Polar Environment Comprehensive Investigation and Assessment Program (CHINARE2014-02-01), and Shanghai Rising-Star Program (11QA1407300).|
The personnel involved in the project:
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.
|Study Extent||Samples were taken in January 2013, during the 29th Chinese National Antarctic Research Expedition at Greatwall Cove and Ardley Cove (king George Island, Antarctica)|
Method step description:
- DNA extraction was performed as described by Luo et al. (2009).
- 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.
- 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).
- 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