characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2014-03-03T16:32:10Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-08T16:54:40Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA2258963
characteristics.characteristic: External Id
characteristics.text: Sapienza University of Rome
characteristics.characteristic: INSDC center name
characteristics.text: 2014-03-03T16:32:10Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-08T16:54:40Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS372690
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-2104:Jurkat_rep1
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: Jurkat
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0002796
characteristics.characteristic: cell line
characteristics.text: human
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9606
characteristics.characteristic: common name
characteristics.text: Protocols: Jurkat cells were grown in RPMI medium supplemented with 10% (v/v) fetal bovine serum, 2mM L-Glutamine and Penicillin-Streptomycin. Nuclei of 350 x 10E+6 Jurkat cells were lysed in IP-buffer supplemented with RNasin (Promega). Nuclear lysates were clarified and pre-cleared in the presence of Protein G-sepharose beads for 2 hr 4 degrees C under constant rotation and then filtered through a 0.45 um filter. Lysate aliquots were taken out as Input for RNA and protein isolation. Antibody anti-AGO2 (11A9, Ascenion) and isotype-matched IgG (mock_IP) were coupled to Protein G-sepharose beads for 2 h at 4 degrees C in IP-buffer containing 1 mg/ml heparin (Sigma). The pre-cleared nuclear lysate was incubated with antibody-coupled beads overnight at 4 degrees C under constant rotation. IP samples were washed once with IP-buffer for 5 min at 4 degrees C, and three times with IP-wash buffer (50 mM Tris-HCl, pH 7.5; 300 mM NaCl; 5 mM MgCl2; and 0.05% Nonidet P-40) at 4 degrees C. An aliquot was taken out for western blot in order to check immunoprecipitation efficiency. For RNA isolation, Input and IP samples were DNAseI- and Proteinase K-treated. Total RNA and co-precipitated RNA was extracted by phenol:chloroform:isopropyl alcohol and precipitated in ethanol. Small RNA libraries were constructed using the TruSeq SmallRNA Sample preparation kit (Illumina)
characteristics.characteristic: description
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: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: frozen specimen
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/OBI_0000922
characteristics.characteristic: specimen with known storage state
characteristics.text: Jurkat_rep1
characteristics.characteristic: title