characteristics.text: 2013-08-30T17:00:54Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-08T16:36:41Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA2149044
characteristics.characteristic: External Id
characteristics.text: College of Life Science, Sun Yat-Sen University
characteristics.characteristic: INSDC center name
characteristics.text: 2013-08-30T17:00:54Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-08T16:36:41Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS322758
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-1748:leucocyte
characteristics.characteristic: Submitter Id
characteristics.text: 40 year
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/UO_0000036
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: Indo-pacific humpbacked dolphin
characteristics.characteristic: common name
characteristics.text: Protocols: The blood sample of the Indo-Pacific humpback dolphin was obtained from a wild male adult individual The fresh blood were collected into EDTA-containing tubes, and then centrifuged for 5 min at 400-500g. The red supernatant was discarded and then 18 ml of Red Blood Cell Lysis Buffer (Beyotime Institute of Biotechnology) were added, mixed gently by flicking the tubes and lasted for 10 min. The tubes were then centrifuged at 1000g for 5 minutes at 4 degrees C to collect the leukocyte pellet. Total RNA was extracted from the leucocytes of Indo-Pacific humpback dolphin using TRIzol reagent (Invitrogen) according to the manufacturer's protocol. Illumina sequencing was performed at Beijing Genomics Institute (BGI)-Shenzhen, China. mRNA with poly(A) tail was isolated from 20ug total RNA treated with DNase I using Magnetic Oligo (dT) Beads (Illumina). The mRNA was fragmented into small pieces (200-700 bp) by treatment with divalent cations at 94 degrees C for 5 minutes. With random hex-amer primers (Illumina), the double-stranded cDNA was synthesized using the SuperScript double-stranded cDNA synthesis kit (Invitrogen) and was further subjected to end-repair using T4 DNA polymerase, the Klenow fragment, and T4 polynucleotide kinase followed by a single A base addition using Klenow 3' to 5' exo-polymerase, then ligated with an adapter or index adapter using T4 DNA ligase. To select the proper templates for downstream enrichment, the products of ligation reaction were purified on 2% agarose gel. The cDNA fragments (about 200 bp) were excised from the gel. Fifteen rounds of PCR amplification were carried out to enrich the purified cDNA template using PCR primer PE 1.0 and 2.0 (Illumina) with phusion DNA polymerase. Finally, the cDNA library was constructed with 200 bp insertion fragment.
characteristics.characteristic: description
characteristics.text: adult
characteristics.characteristic: developmental stage
characteristics.text: Sousa chinensis
characteristics.characteristic: organism
characteristics.text: Sousa chinensis
characteristics.characteristic: organism
characteristics.text: Leucocyte
characteristics.characteristic: organism part
characteristics.text: Sousa chinensis
characteristics.characteristic: scientific_name
characteristics.text: male
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/PATO_0000384
characteristics.characteristic: sex
characteristics.text: fresh specimen
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/OBI_0000971
characteristics.characteristic: specimen with known storage state
characteristics.text: leucocyte
characteristics.characteristic: title