Contributors | Affiliation | Role |
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Goetze, Erica | University of Hawaiʻi at Mānoa (SOEST) | Principal Investigator |
Copley, Nancy | Woods Hole Oceanographic Institution (WHOI BCO-DMO) | BCO-DMO Data Manager |
This dataset includes RADSeq data as well as NCBI Short Read Archive (SRA) BioProject and BioSample accessions and collection metadata from animals collected on Atlantic Meridional Transect 22 (AMT22) in Oct. - Nov. 2012. Field work was conducted on the RRS James Cook cruise JC079. See NCBI GenBank Bioproject PRJNA369277 [https://www.ncbi.nlm.nih.gov/bioproject/PRJNA369277]
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Please refer to the paper for methodological details. If you have further questions, please contact the corresponding author (Dr. Erica Goetze): egoetze[at]hawaii[dot]edu.
From the cruise report:
Sample collection. Plankton samples were collected with 0.71m diameter bongo nets (200, 333 µm), and with an RMT1 midwater trawl (333 µm) that has a nominal mouth area of 1m2. A total of 50 plankton tows were conducted along the cruise leg (Table 1), with 35 tows conducted using the bongo and 14 samples collected with the RMT net. The bongo tows were oblique tows that sampled from between 211 to 488 m depth and the surface (324m average maximum depth of tow). The bongo samples will be used for quantitative estimates of animal abundance along the cruise leg (target species only, tows conducted with timedepth-recorder and flowmeter). The RMT tows were also oblique tows that sampled between 62 to 216 m depth and the surface (153 m average maximum depth of tow). All tows except one (station 42) were conducted at night, in order to efficiently sample the migratory community.
Sample handling and preservation. All plankton from the 200 µm mesh bongo net was preserved immediately in 100% ethyl alcohol for use in molecular studies, including DNA sequencing and microsatellite genotyping (and possibly RAD tag sequencing), in addition to estimates of abundance of target species. Plankton material from the 333 µm mesh bongo net and the RMT net was sorted live immediately following collection, and animals were individually identified, and preserved in acetone, RNALater, cryopreserved, and in some cases used for live imaging prior to preservation. These animals will be used for molecular, genomic and transcriptomic analyses. Both RNA/DNA ratios and prosome length - dry weight relationships will be used as measures of animal condition in copepods. In total, over 17,000 animals from 40 target species were individually sorted and preserved for this panel of measurements. Following live sorting and imaging of the 333 µm samples, the remaining plankton was preserved either in 4% buffered formalin or 100% ethyl alcohol for morphological studies.
Illumina sequence files:
This submission consists of reduced representation genomic data for 26 individual pteropod specimens in the genus Cuvierina. Specimens were collected during the Atlantic Meridional Transect cruise in 2012 (AMT22) as well as a Mar-Eco cruise. Our research goals were to examine genomic divergence and species boundaries in these morphologically and ecologically distinct pteropod populations in adjacent ocean regions of the subtropical and equatorial Atlantic. We are interested in local adaptation of these populations to distinct oceanographic habitats.
Genetic data consists of 300 bp paired-end sequence reads generated using an Illumina MiSeq sequencer. Two MiSeq lanes were run; approximately 400mB to 1GB of data were generated for each library, with a mean of 700mB per individual. Libraries for individuals #1, 2, 3, 4, 9, 10, 11, 12, 16, 17, 18, 19, and 25 were run together on a single MiSeq lane, and libraries for individuals #5, 6, 7, 8, 13, 14, 15, 20, 21, 22, 23, 24, and 26 were run together on a second lane.
The raw data was cleaned of adapters, trimmed of poor sequence, and filtered for non-specific sequencing. Cutadapt v. 1.9.1 was used to remove the Truseq Illumina Adapters (LT), using a 90% mismatch parameter (-e 0.10) and a minimum adapter match overlap of 12 bp (-O 12). Reads that did not contain an adapter (potentially non-specific sequencing) were removed from the dataset (--discard-trimmed). Read 1 and read 2 were paired using the custom python script, "fastqCombinePairedEnd.py", and read pairs were verified using the custom Perl script, "FastqPairedEndValidator.pl". Trimmed, cleaned, and paired reads were checked for sequence quality using FastQC.
This post-sequence processing resulted in three files included for each individual library: *.R1.fq (All read 1s that paired with a read 2), *.R2.fq (All read 2s that paired with a read 1), and *.R1.clean.fq_singles.fastq (unpaired reads), with the * representing each respective putative Cuvierina sp. individual.
Contact: Erica Goetze for any questions, or for subsequent use of these data.
BCO-DMO Processing Notes:
added conventional header with dataset name, PI name, version date
modified parameter names to conform with BCO-DMO naming conventions
combined SRA metadata with collection information
converted latitude and longitude to decimal degrees
Parameter | Description | Units |
biosample_accession | NCBI BioSample accession number | unitless |
library_ID | NCBI Library identifier | unitless |
filename | NCBI filename | unitless |
sample_title | NCBI sample title | unitless |
cruise_id | cruise identifier | unitless |
station | station number | unitless |
date_collection | collection date formatted as yyyy-mm-dd | unitless |
lat_collection | latitude; north is positive | decimal degrees |
lon_collection | longitude; east is positive | decimal degrees |
sample_id | sample identifier | unitless |
sample_name | short sample identifier | unitless |
Dataset-specific Instrument Name | Illumina HiSeq 2500 |
Generic Instrument Name | Automated DNA Sequencer |
Generic Instrument Description | General term for a laboratory instrument used for deciphering the order of bases in a strand of DNA. Sanger sequencers detect fluorescence from different dyes that are used to identify the A, C, G, and T extension reactions. Contemporary or Pyrosequencer methods are based on detecting the activity of DNA polymerase (a DNA synthesizing enzyme) with another chemoluminescent enzyme. Essentially, the method allows sequencing of a single strand of DNA by synthesizing the complementary strand along it, one base pair at a time, and detecting which base was actually added at each step. |
Dataset-specific Instrument Name | |
Generic Instrument Name | Thermal Cycler |
Generic Instrument Description | A thermal cycler or "thermocycler" is a general term for a type of laboratory apparatus, commonly used for performing polymerase chain reaction (PCR), that is capable of repeatedly altering and maintaining specific temperatures for defined periods of time. The device has a thermal block with holes where tubes with the PCR reaction mixtures can be inserted. The cycler then raises and lowers the temperature of the block in discrete, pre-programmed steps. They can also be used to facilitate other temperature-sensitive reactions, including restriction enzyme digestion or rapid diagnostics.
(adapted from http://serc.carleton.edu/microbelife/research_methods/genomics/pcr.html) |
Website | |
Platform | RRS James Cook |
Report | |
Start Date | 2012-10-10 |
End Date | 2012-11-24 |
Description | The AMT22 cruise set sail from Southampton in the UK on 10 October 2012 and arrived in Punta Arenas, Chile on 24 November 2012.
The final cruise report and other cruise information, including all science components, can be found online at the Atlantic Meridional Transect webpage (http://www.amt-uk.org/Cruises), or through the British Oceanographic Data Centre (BODC) (http://www.bodc.ac.uk/projects/uk/amt/).
Zooplankton ecology data from the project "Does habitat specialization drive population genetic structure of oceanic zooplankton?" (NSF OCE-1029478) were collected on this cruise. |
Description from NSF award abstract:
Marine zooplankton show strong ecological responses to climate change, but little is known about their capacity for evolutionary response. Many authors have assumed that the evolutionary potential of zooplankton is limited. However, recent studies provide circumstantial evidence for the idea that selection is a dominant evolutionary force acting on these species, and that genetic isolation can be achieved at regional spatial scales in pelagic habitats. This RAPID project will take advantage of a unique opportunity for basin-scale transect sampling through participation in the Atlantic Meridional Transect (AMT) cruise in 2014. The cruise will traverse more than 90 degrees of latitude in the Atlantic Ocean and include boreal-temperate, subtropical and tropical waters. Zooplankton samples will be collected along the transect, and mitochondrial and microsatellite markers will be used to identify the geographic location of strong genetic breaks within three copepod species. Bayesian and coalescent analytical techniques will test if these regions act as dispersal barriers. The physiological condition of animals collected in distinct ocean habitats will be assessed by measurements of egg production (at sea) as well as body size (condition index), dry weight, and carbon and nitrogen content. The PI will test the prediction that ocean regions that serve as dispersal barriers for marine holoplankton are areas of poor-quality habitat for the target species, and that this is a dominant mechanism driving population genetic structure in oceanic zooplankton.
Note: This project is funded by an NSF RAPID award. This RAPID grant supported the shiptime costs, and all the sampling reported in the AMT24 zooplankton ecology cruise report (PDF).
Online science outreach blog at: https://atlanticplankton.wordpress.com
Funding Source | Award |
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NSF Division of Ocean Sciences (NSF OCE) |