Solamente metadatos

Airborne Bacteria from Miers Valley, Antarctica

Última versión Publicado por SCAR - Microbial Antarctic Resource System en 19 de marzo de 2019 SCAR - Microbial Antarctic Resource System
Fecha de publicación:
19 de marzo de 2019
Licencia:
CC-BY 4.0

Descargue la última versión de los metadatos como EML o RTF:

Metadatos como un archivo EML descargar en Inglés (10 KB)
Metadatos como un archivo RTF descargar en Inglés (12 KB)

Descripción

Amplicon dataset of bacteria captured 1 m above the ground in two locations in Miers Valley (Antarctica).

Versiones

La siguiente tabla muestra sólo las versiones publicadas del recurso que son de acceso público.

¿Cómo referenciar?

Los usuarios deben citar este trabajo de la siguiente manera:

Bottos E, Woo A, Zawar-Reza P, Pointing S, Cary C (2018): Airborne Bacteria from Miers Valley, Antarctica. v1.2. SCAR - Microbial Antarctic Resource System. Dataset/Metadata. https://ipt.biodiversity.aq/resource?r=airborne_bacteria_miers_antarctica&v=1.2

Derechos

Los usuarios deben respetar los siguientes derechos de uso:

El publicador y propietario de los derechos de este trabajo es SCAR - Microbial Antarctic Resource System. Este trabajo está autorizado bajo una Licencia Creative Commons Atribución/Reconocimiento 4.0 Internacional (CC-BY) 4.0.

Registro GBIF

Este recurso ha sido registrado en GBIF con el siguiente UUID: 6a1177d0-ce25-4611-9666-a3d58cdea8c5.  SCAR - Microbial Antarctic Resource System publica este recurso y está registrado en GBIF como un publicador de datos avalado por Scientific Committee on Antarctic Research.

Palabras clave

Metadata

Contactos

¿Quién creó el recurso?:
-

Eric Bottos
University of Waikato
Hamilton
NZ
Antony Woo
Sorbonne Paris Cité
Paris
FR
Peyman Zawar-Reza
University of Waikato
Hamilton
NZ
Stephen Pointing
University of Waikato
Hamilton
NZ
Craig Cary
University of Waikato
Hamilton
NZ

¿Quién puede resolver dudas acerca del recurso?:

Stephen Pointing
University of Waikato
Hamilton
NZ

¿Quién documentó los metadatos?:
-

Maxime Sweetlove
Research assistent
Royal Belgian Institute for Natural Sciences
Brussels
BE

¿Quién más está asociado con el recurso?:

Cobertura geográfica

Miers Valley, Antarctica

Coordenadas límite Latitud Mínima Longitud Mínima [-78,114, 163,786], Latitud Máxima Longitud Máxima [-78,096, 163,787]

Cobertura taxonómica

16S ssu rRNA

Dominio Bacteria (Bacteria)

Cobertura temporal

Periodo de formación 2009-12-11 2010-01-25

Métodos de muestreo

Aerosol samples were collected by filtration onto 0.2-μm-pore-size polycarbonate filters by impaction using solar-powered pumps (SKC, 224-PCXR8, Eighty Four, PA, USA) mounted 1 m above the ground in each location. Filters inserted into cassette apparatus but not exposed to air flow were used as controls. All filters and cassettes were UV-sterilized and rinsed with 70 % alcohol before use. Sampling apparatus was deployed 1 m above the ground surrounded with a 2 mm gauze baffle on the Miers Valley Floor (78°05′.78S, 163°47′.25E, approx. 270 m) and Miers Valley Ridge (78°06′.83S, 163°47′.18E, approx. 550 m).

Área de Estudio Samples were collected from a continuous filtration period December 11, 2009 to January 25, 2010 (55 days), with an estimated sample volume of 75,000 l for each location.

Descripción de la metodología paso a paso:

  1. Air sampling (see sampling description)
  2. Filters were stored at −20 °C during transit from Antarctica and until processed.
  3. Total DNA was extracted directly from the filters using the DNeasy Plant Mini Kit (Qiagen, CA, USA), after first washing with kit lysis buffer for 10 min. The remaining steps of the extraction were carried out according to the manufacturers instructions. Recovered DNA was quantified using NanoDrop™ (Thermo Scientific, Waltham, MA, USA).
  4. For each sample, PCR targeting the V5–V7 region of the 16S rRNA gene was completed in duplicate. Each 30 μl reaction contained 1× PrimeSTAR buffer, 0.2 mM dNTPs, 0.75 U PrimeSTAR HS DNA Polymerase (Takara Holdings, Kyoto, Japan), 0.4 μM of primers Tx9 (5′-GGATTAGAWACCCBGGTAGTC-3′) and 1391R (5′-GACGGGCRGTGWGTRCA-3′), and 5 μl of template DNA. Thermal cycling conditions consisted of 94 °C for 3 min; 30 cycles of 94 °C for 20 s, 52 °C for 20 s, and 72 °C for 45 s; and 72 °C for 3 min. All thermal cycling was completed on a Bio-Rad DNA Engine Peltier Thermal Cycler 200 (Bio-Rad, Hercules, CA, USA). Duplicate reactions were pooled, and amplicons were size-selected from agarose gels using a MO BIO Gel Extraction Kit (MO BIO Laboratories, Carlsbad, CA, USA). Gel extracted products were cleaned using an Agencourt AMPure XP system (Beckman Coulter, Brea, CA, USA) and quantified using a Qubit dsDNA HS Assay Kit and Qubit 2.0 Fluorometer (Life Technologies, Carlsbad, CA, USA).
  5. To prepare the amplicons for sequencing, a second round of PCR was completed in triplicate. PCR reactions were prepared as outlined above, but using 10 ng of purified amplicon as the template and primers MIDX-Tx9F (5′-CCATCTCATCCCTGCGTGTCTCCGACTCAG-MID-GGATTAGAWACCCBGGTAGTC-3′) and BacB_1391R (5′-CCTATCCCCTGTGTGCCTTGGCAGTCTCAG-GACGGGCRGTGWGTRCA-3′), adapted for one-way reads with unique MID identifiers for each sample. Thermal cycling conditions were as outlined above, but reduced to 13 cycles. Triplicate products were pooled, gel-extracted, cleaned, and quantified as outlined above, before quantification of amplifiable molecules using a KAPA Library Quantification Kit for Roche 454 Titanium/Universal (Kapa Biosystems, Woburn, MA, USA) on a Corbett Rotor-Gene 6000 real-time thermal cycler (Life Technologies). Amplicons were sequenced using the GS Junior Titanium emPCR Kit (Lib-L), the GS Junior Titanium Sequencing Kit, PicoTitrePlate Kit, and GS Junior System (Roche 454 Life Sciences, Branford, CT, USA) at The University of Waikato DNA Sequencing Facility.

Referencias bibliográficas

  1. Bottos, E. M., Woo, A. C., Zawar-Reza, P., Pointing, S. B., & Cary, S. C. (2014). Airborne bacterial populations above desert soils of the McMurdo Dry Valleys, Antarctica. Microbial ecology, 67(1), 120-128.

Metadatos adicionales

Identificadores alternativos 6a1177d0-ce25-4611-9666-a3d58cdea8c5
https://ipt.biodiversity.aq/resource?r=airborne_bacteria_miers_antarctica