|Harvell, Drew||Cornell University (Cornell)||Principal Investigator|
|Mydlarz, Laura||University of Texas at Arlington (UT Arlington)||Co-Principal Investigator|
|Burge, Colleen||Cornell University (Cornell)||Contact|
|Rauch, Shannon||Woods Hole Oceanographic Institution (WHOI BCO-DMO)||BCO-DMO Data Manager|
Accession numbers to NCBI's GenBank are provided for Labyrinthulomycetes detected in sea fans collected in Florida and Puerto Rico.
Burge CA, Douglas N, Conti-Jerpe I, Weil E, Roberts S, Friedman CS & CD Harvell. In press (May 2012) Friend or foe: the association of Labyrinthulomycetes with the Caribbean sea fan, Gorgonia ventalina. Dis Aquat Org.
Sampling and Analytical Methodology:
Healthy and diseased sea fans were collected from Florida and Puerto Rico. Labyrinthulomycetes cells were isolated from a diseased sea fan from Florida; DNA extracted, amplified, cloned, sequenced, and deposited with accession # JQ248602. Fragments of healthy and diseased sea fans were extracted, amplified, cloned and sequenced under accession numbers JQ248603 (Florida), JQ248604 (Florida), JQ248605 (Puerto Rico), and JQ248606 (Puerto Rico).
Labyrinthulomycota small-subunit ribosomal DNA (SSU rRNA) specific primers, Laby-A and Laby-Y, were used (Stokes et al., 2002). Those containing bands of the expected (~ 430 bp) size were purified using the QIAquick PCR kit (Qiagen,) and sent to the Cornell University Life Sciences Core Laboratories for direct sequencing on an Applied Biosystems Automated 3730 DNA Analyzer. Although direct sequencing of all of the PCR products showed unambiguous DNA chromatographs throughout, the homogeneity of samples was additionally confirmed by cloning the PCR products into the TOPO-TA vector, using the TOPO TA cloning kit (Invitrogen) for sequencing of 2 to 5 clones of each isolate.
The BLAST algorithm (Altschul et al.1990) was used to compare resulting sequences (overlapping 398 nt from Florida samples and 292 nt from Puerto Rico samples) with those deposited in the National Center for Biotechnology Information (NCBI) GenBank database, and alignments were made using the EMBL-EBI Clustal W tool (Larkin et al. 2007, Goujan et al. 2010). Representative sequences from the cultured microorganism and sea fan isolates were submitted to GenBank.
|site||Name of the reef where the sample was collected.||text|
|lat||Latitude of the collection site. North = positive.||decimal degrees|
|lon||Longitude of the collection site. West = negative.||decimal degrees|
|sequence_name||Name of the sequence/sample.||text|
|accession_number||Assigned accession number and link to GenBank.||unitless|
|Dataset-specific Instrument Name|| |
Automated DNA Sequencer
|Generic Instrument Name|| |
Automated DNA Sequencer
|Dataset-specific Description|| |
Direct sequencing was performed at the Cornell University Life Sciences Core Laboratories on an Applied Biosystems Automated 3730 DNA Analyzer
|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.
|Start Date|| |
|End Date|| |
Four dive sites for the Harvell/Mydlarz project 'Influence of Temperature and Acidification on the Dynamics of Coral Co-Infection and Resistance': Big Pine Ledges, Florida Keys: 24° 33.207 N, 81° 22.731 W Laurel patch reef, La Parguera, Puerto Rico: 17° 56.608 N, 67° 03.208 W Media Luna, La Parguera, Puerto Rico: 17°56.093 N, 67°02.931 W (3 to 18 m depths) Buoy, La Parguera, Puerto Rico: 17° 53.38 N, 66° 59.09 W (18 to 25 m depths)
This document is created by info v 4.1f 5 Oct 2018 from the content of the BCO-DMO metadata database. 2020-02-18 13:24:34