Arctic multi-year sea ice Bacteria

版本

以下的表格只顯示可公開存取資源的已發布版本。

如何引用

研究者應依照以下指示引用此資源。:

Hatam I, Lange B, Beckers J, Haas C, Lanoil B, Charchuk R (2019): Arctic multi-year sea ice Bacteria. v1.2. SCAR - Microbial Antarctic Resource System. Dataset/Metadata. https://ipt.biodiversity.aq/resource?r=arctic_multiyear_sea_ice_bacteria&v=1.2

權利

研究者應尊重以下權利聲明。:

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.

GBIF 註冊

此資源已向GBIF註冊，並指定以下之GBIF UUID: 9f8879f7-ae24-4c6c-9209-d62408e5dcbb。  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

Landfast ice off the northern shore of northern Ellesmere Island Nunavut, Canada (82.54905°N, −62.37685°W).

分類群涵蓋範圍

Bacteria, targeted with the 16S ssu rRNA marker gene (v1-v3 region)

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

Two parallel cores were sampled using Kovacs Mark II 9 cm corer (Kovacs Enterprise, Roseburg, Oregon) and a 36 V electric hand drill. Prior to drilling, the core barrel was rinsed with sterile deionized water (Milli-Q Integral Water Purification System, EMD Millipore Corporation, Billerica, MA). The two cores were immediately sectioned on site to 30-cm intervals using a hand saw (rinsed in deionized water and wiped with an ethanol wipe) and placed in UV-sterilized polypropylene bags.

方法步驟描述:

1. Ice samples used were thawed at room temperature in the dark, and were filtered individually through 0.22-μm-pore-size polyethersulfone membrane filters (Pall, Mississauga, ON, Canada). Direct melting was used to avoid nonspecific addition of DNA and dilution of samples. All glassware was sterilized using 10% household bleach solution followed by thorough washing in sterile deionized water between samples to prevent cross-contamination. Each filter was placed in a microfuge tube and submerged in RNAlater® solution (Life Technologies, Burlington, ON, Canada) and stored at −20 °C for later DNA extraction.
2. DNA was extracted from preserved filters by bead beating using the FastDNA® SPIN Kit for Soil (MP Biomedicals, Solon, OH) as instructed by the manufacturer. Filters from equivalent 30-cm sections of duplicate ice cores were combined prior to DNA extraction to ensure sufficient DNA yield. Seawater duplicate samples were processed individually.
3. Molecular Research LP (Shallowater, TX) performed pyrosequencing of the V1-V3 regions of the bacterial 16S rRNA gene as described in Dowd et al. (2008). Amplification was performed using HotStarTaq Plus Master Mix Kit (Qiagen, Valencia, CA) under the following conditions: 94 °C for 3 min, followed by 28 cycles of 94 °C for 30 s; 53 °C for 40 s; and 72 °C for 1 min with a final elongation step at 72 °C for 5 min. Gene-specific forward PCR primer sequences were tagged with the sequencing adapters for GS FLX Titanium chemistry, an 8 base barcode, and a linker sequence. All PCRs were performed in triplicate, mixed in equal concentrations, and purified using Agencourt AMPure beads (Agencourt Bioscience Corporation, MA). FLX-Titanium amplicon pyrosequencing was performed using the Genome Sequencer FLX System (Roche, Branford, CT).

引用文獻

1. Hatam, I., Charchuk, R., Lange, B., Beckers, J., Haas, C., & Lanoil, B. (2014). Distinct bacterial assemblages reside at different depths in Arctic multiyear sea ice. FEMS microbiology ecology, 90(1), 115-125. https://doi.org/10.1111/1574-6941.12377

額外的詮釋資料