characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2014-05-01T17:00:44Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2014-03-24T14:08:40Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA2422316
characteristics.characteristic: External Id
characteristics.text: Sars International Centre for Marine Molecular Biology
characteristics.characteristic: INSDC center name
characteristics.text: 2014-05-01T17:00:44Z
characteristics.characteristic: INSDC first public
characteristics.text: 2014-03-24T14:08:40Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS427223
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-2431:late_cleavage_replica2
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: Protocols: Adult Sycon ciliatum specimens were collected from fjords located near Bergen, Norway, and transported to laboratory in sea water collected at the place of their growth, with temperature between 6 and 16 degrees Celsius depending on time of the year. They were gradually (1-3 hours) acclimatized to ambient temperature, avoiding temperatures exceeding 20 degrees. Field-collected water was gradually exchanged for sterile filtered natural sea water by replacing 1/4th to 1/3rd parts with the filtered water every 20-30 minutes 3-4 times. With aid of dissecting microscope, the samples were separated from the algae and cleaned from macroscopic contaminations by gently blowing water at them.After several water changes, the adults were cut transversely so that the top and bottom parts were discarded, and only mid-body fragments (contining oocytes or embryos) were selected for RNA isolation. To obtain larvae, sponges containing larvae were kept in plastic cups with a pellet of 1-Hexadecanol (to avoid trapping and damage of the larvae by surface tension) until the larvae were released (1-4days). The larvae swimming near the surface were poured into clean Petri dishes. For RNA isolation from larvae, larvae were collected by glass pasteur pipet with heat-polished tip or using automatic P200 pipette with a standard tip into 1.5 ml tubes and further concentrated by centrifugation. For nucleic acids isolation from juveniles, the larvae were kept in the petri dishes, including a pellet of 1-Hexadecanol, until settlement and metamorphosis, which took 6-10 days to complete, and which was periodically monitored under dissecting microscope. None. Total RNA and DNA were isolated using the AllPrep DNA/RNA Mini Kit (Qiagen) according to manufacturerﾒs instructions. Adult tissue pieces were snap-frozen in liquid nitrogen and processed immediately or stored at -80 degrees until processing. In case of larvae, water was removed after centrifugation and replaced with the lysis buffer, and sample disrupted in 1.5 ml tubes in the buffer using a micropestle. In case of juveniles, which were attached to the bottom of the petri dishes, water was replaced with the lysis buffer and juveniles scraped of the bottom using cell culture scraper before transferring to 1.5 ml tubes and disruption. Isolated RNA and DNA were analysed on agarose gels and quantified by Nanodrop, some of the RNA samples were additionally analysed using Bioanalyzer. The libraries were prepared according to Illumina instructions for RNA sample preparation "TruSeq RNA V2".
characteristics.characteristic: description
characteristics.text: late cleavage
characteristics.characteristic: developmental stage
characteristics.text: Sycon ciliatum
characteristics.characteristic: organism
characteristics.text: Sycon ciliatum
characteristics.characteristic: organism
characteristics.text: Sycon ciliatum
characteristics.characteristic: scientific_name
characteristics.text: fresh
characteristics.characteristic: specimen with known storage state
characteristics.text: Sycon ciliatum
characteristics.characteristic: strain
characteristics.text: late_cleavage_replica2
characteristics.characteristic: title