characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2018-05-16T17:03:07Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-16T15:14:57Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA104700134
characteristics.characteristic: External Id
characteristics.text: Temple University Lewis Katz School of Medicine
characteristics.characteristic: INSDC center name
characteristics.text: 2018-05-16T17:03:07Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-16T15:14:57Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS2301517
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-6604:LPC
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: endothelial cell
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/CL_0000115
characteristics.characteristic: cell type
characteristics.text: human
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9606
characteristics.characteristic: common name
characteristics.text: Protocols: HAECs were collected using RNAeasy kit. Human aortic endothelial cells (HAECs) (Lonza, CC2535; Walkersville, MD) were cultured in medium M199 (Hyclone laboratories, Logan, UT) supplemented with 15% fetal bovine serum (FBS; HyClone); endothelial cell growth supplement (ECGS, 50μg /mL; BD Biosciences, San Jose, CA); heparin (50μg/mL); and 1% penicillin, streptomycin, and amphotericin (PSA; Invitrogen, Carlsbad, CA). HAECs were grown on 0.2% gelatin-coated flasks, plates, or dishes and experiments were performed at passage 9. 18-hour vehicle or lysophosphatidylcholine (16:0) treatment RNA is extracted using RNeasy kit, then mRNA and non-coding RNAs are enriched by removing rRNA from the total RNA with kit. By using the fragmentation buffer, the mRNAs are fragmented into short fragments (about 200~500nt), then the first-strand cDNA is synthesized by random hexamer-primer using the fragments as templates, and dTTP is substituted by dUTP during the synthesis of the second strand. Short fragments are purified and resolved with elution buffer for end reparation and single nucleotide A (adenine) addition. After that, the short fragments are connected with adapters, then the second strand is degraded finally using UNG(Uracil-N-Glycosylase). After agarose gel electrophoresis, the suitable fragments are selected for the PCR amplification as templates. During the quality control steps, Agilent 2100 Bioanaylzer and ABI StepOnePlus Real-Time PCR System are used in quantification and qualification of the sample library.
characteristics.characteristic: description
characteristics.text: wild type genotype
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0005168
characteristics.characteristic: genotype
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9606
characteristics.characteristic: organism
characteristics.text: aorta
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/UBERON_0000947
characteristics.characteristic: organism part
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: LPC
characteristics.characteristic: title