characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2015-02-02T17:02:27Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-08T22:08:36Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA3171408
characteristics.characteristic: External Id
characteristics.text: Programme Carte d'Identitite des Tumeurs
characteristics.characteristic: INSDC center name
characteristics.text: 2015-02-02T17:02:27Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-08T22:08:36Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS625799
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-2833:Sample 56
characteristics.characteristic: Submitter Id
characteristics.text: 61 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: human
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9606
characteristics.characteristic: common name
characteristics.text: Protocols: miRNA-Seq libraries are performed from at least 1ug of extracted total RNA with a RIN greater than 7. Before starting, total RNAs are purified with miRNeasy kit wich allows the selection of the small RNA fraction less than 100b. first, a 3' adenylated DNA adaptor is ligated to the enriched sample in the absence of ATP preventing the self-ligation of miRNAs.Then a 5' RNA adaptor is ligated in the presence of ATP at the other end of the miRNAs.The RT primer complementary of the 3' adaptor is added at this stage with which it will form a duplex thereby reducing the ligation between adaptors.A reverse transcription is therefore perfomed from the RT primer and finally these captured miRNAs are amplified by PCR with primers complementary to the 3' and 5' adaptors.During this PCR a specific barcode is incorporated allowing individualisation of each library.Each PCR is loaded on the Fragment Analyzer (AATI) for a precise quantification of each miRNA peak of interest.Based on these results a equimolar pool of about ten of different samples are performed.Finally the pooled PCR product is loaded on PAGE in order to excise the band of miRNA that is extracted and purified on a Qiagen MinElute column.Finally the pooled PCR product is loaded on PAGE in order to excise the band of miRNA that is extracted and purified on a Qiagen MinElute column.
characteristics.characteristic: description
characteristics.text: Pheochromocytoma
characteristics.characteristic: disease
characteristics.text: HS_181
characteristics.characteristic: individual
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: 26 month
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/UO_0000035
characteristics.characteristic: overall survival delay
characteristics.text: 0
characteristics.characteristic: overall survival event
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: female
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/PATO_0000383
characteristics.characteristic: sex
characteristics.text: Sample 56
characteristics.characteristic: title