Amplicon dataset of Bacteria (16S ssu rRNA) living around mummified seal carcasses in the Antarctic Dry Valleys
Tiao G, Lee C, McDonald I, Cowan D, Cary C (2019): Bacteria on mummified seals in the Antarctic Dry Valleys. v1.2. SCAR - Microbial Antarctic Resource System. Dataset/Metadata. https://ipt.biodiversity.aq/resource?r=antarctic_dry_valley_mummified_seal_microbiome&v=1.2
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.
soil samples from under and around mummified crabeater seal near Lake Miers (S 78°05′36″, E 163°51′22″), Miers Valley, Victoria Land, Antarctica, that was transported to a similar site 20 m north.
|界定座標範圍||緯度南界 經度西界 [-78.933, 163.856], 緯度北界 經度東界 [-78.933, 163.856]|
Amplicin sequencing of Bacteria (16S ssu rRNA gene)
|起始日期 / 結束日期||2005-01-15 / 2009-01-15|
|經費來源||This work was supported through grants from the New Zealand Foundation for Research Science and Technology, Antarctica New Zealand, and the National Science Foundation (OPP-0739648 and 0944560). Additional support was provided by a George Peabody Gardner Fellowship, by the New Zealand Foundation for Research, Science and Technology Postdoctoral Research Fellowship and the New Zealand Marsden Foundation (UOW1003).|
Soil samples (>10 g each) were collected aseptically at a depth of 0.05m. Samples were stored in sterile plastic containers at −80 °C until analysis.
|研究範圍||On 30 January 2006, a mummified crabeater seal was carried from its original resting site near Lake Miers (S 78°05′36″, E 163°51′22″), Miers Valley, Victoria Land, Antarctica, to a new site of similar geomorphology 20 m north. Samples for amplicon sequencing were taken from the original site before transportation (austral summer 2005), the new site before transportation (control, austral summer 2006), and the new site after incubation (austral summer 2009)|
- 16S rRNA gene were generated using the primer pair Tx9 (5′-GGATTAGAWACCCBGGTAGTC-3′) and 1391R (5′-GACGGGCRGTGWGTRCA-3′). PCR was performed in triplicate and pooled to reduce stochastic variability between reactions. Each 30 μl reaction contained 5 or 20 ng of extracted community DNA, Pfx polymerase and platinum polymerase (0.5 U each; Invitrogen), 1× Pfx PCR buffer with Pfx enhancer, 0.2 mM dNTPs, 1 mM MgCl2, 0.02 mg ml−1 BSA, 0.8 μM of forward and reverse primer, and PCR-grade water. Thermal cycling conditions were 94 °C for 2 min; 18 or 24 cycles of 94 °C for 15 s, 55 °C for 30 s and 68 °C for 1 min; and 68 °C for 3 min.
- Amplicons were size-selected and purified using polyacrylamide gel electrophoresis before being prepared for pyrosequencing by Taxon Biosciences (Tiburon, CA, USA).
- Tiao, G., Lee, C. K., McDonald, I. R., Cowan, D. A., & Cary, S. C. (2012). Rapid microbial response to the presence of an ancient relic in the Antarctic Dry Valleys. Nature Communications, 3, 660.