characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2017-08-07T17:02:55Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2017-03-22T11:23:49Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA103934102
characteristics.characteristic: External Id
characteristics.text: AG Clayton Zentrum fur Molekulare Biologie der Universitat Heidelberg
characteristics.characteristic: INSDC center name
characteristics.text: 2017-08-07T17:02:55Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-03-22T11:23:49Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS1623270
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-5602:PC_2098_Uninduced_2
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: pHD1313/2098
characteristics.characteristic: cell line
characteristics.text: Protocols: Procyclic cells (EATRO 1125 strain) with a Tet-repressor construct (pHD1313) and an inducible RBP10-myc construct (pHD2098) was used. The cells were grown at 27oC in MEM-Pros medium supplemented with 10% heat inactivated fetal bovine serum. To convert procyclic (PC) to bloodstream (BS) forms, RBP10-myc was induced for 48 hours using tetracycline (200ng/ml), the cells were pelleted, resuspended (2x105 cells/ml) in HMI-9 medium and then incubated at 37oC with 5% CO2. For RNA-seq, RNA was isolated from two biological replicates after RBP10 expression for 24h and 48h; uninduced procyclic cells served as control. Two additional RNA samples from re-differentiated bloodstream forms (newly converted BS) and starting EATRO 1125 bloodstream (BS AnTat1.1_2098) cells were included. Trifast reagent (Peqlab, GMBH) was used for total RNA isolation. 5µg of total RNA was depleted of ribosomal RNA (rRNAs) using RNAse H (5U) in presence of 50-base DNA oligos complementary to trypanosome rRNAs. After DNAse I treatment, RNA was purifed using RNA Clean & Concentrator kit (Zymo Reseach, USA). RNA-seq was done at the CellNetworks Deep Sequencing Core Facility at the University of Heidelberg. For library preparation, NEBNext Ultra RNA Library Prep Kit for Illumina (New England BioLabs Inc.) was used according to the manufacturer’s instructions.
characteristics.characteristic: description
characteristics.text: procyclic
characteristics.characteristic: developmental stage
characteristics.text: Trypanosoma brucei
characteristics.characteristic: organism
characteristics.text: Trypanosoma brucei
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_5691
characteristics.characteristic: organism
characteristics.text: Trypanosoma brucei
characteristics.characteristic: scientific_name
characteristics.text: EATRO1125
characteristics.characteristic: strain
characteristics.text: PC_2098_Uninduced_2
characteristics.characteristic: title