characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2018-01-30T17:02:44Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2017-03-13T15:51:25Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA103910555
characteristics.characteristic: External Id
characteristics.text: School of Medicine, Health Science Center, Shenzhen University, Shenzhen, China
characteristics.characteristic: INSDC center name
characteristics.text: 2018-01-30T17:02:44Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-03-13T15:51:25Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS1599723
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-5566:HCT1116-shPBRM1 rep1
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: HCT116
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0002824
characteristics.characteristic: cell line
characteristics.text: human
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9606
characteristics.characteristic: common name
characteristics.text: Protocols: HCT116 cells were infected with lentivirus containing non-targeting shRNA or shRNA targeting PBRM1. After 72h of infection, cells were harvested and total RNA was extracted. Alternatively, HCT116 cells were transfected with non-targeting siRNA or siRNA targeting PBRM1. After 72h of transfection, cells were harvested and total RNA was extracted. Total RNA was extracted from HCT116 cells using RNeasy kit from Qiagen according to manufacturer's instructions Oligo (dT) magnetic beads are used to select mRNA with polyA tail. Fragment the target RNA and reverse transcription to double-strand cDNA (dscDNA) by N6 random primer. End repair the dscDNA with phosphate at 5' end and stickiness 'A' at 3' end, then ligate adaptor with stickiness 'T' at 3' end to the dscDNA. Two specific primers are used to amplify the ligation product. Denature the PCR product by heat and the single strand DNA is cyclized by splint oligo and DNA ligase.
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: HCT1116-shPBRM1 rep1
characteristics.characteristic: title