2024-03-08
ENA first public
ERC000011
ENA-CHECKLIST
Head of Drug Discovery and Evaluation Charles University in Prague
INSDC center name
public
INSDC status
ERS18345606
SRA accession
E-MTAB-13844_2:PH39
Submitter Id
ArrayExpress
broker name
not collected
collection date
Protocols: RNA library screening was carried out as previously described (PMID: 25502887).RITseq library cells were expanded to 2x107 cells in 150 ml HMI-9 and induced with 1 µg/ml of tetracyclin for 24 h prior to PHT-39 addition. PHT-39 was added to the cells at 1 x EC50 concentrations, and the number of viable cells was monitored by daily hemocytometer counting. Cells were harvested at a density of >10e8 and genomic DNA isolated. T. brucei brucei (strain 427 Lister) blood stream form (BSF) parasites were passaged in complete HMI-9 media supplemented with 10% FCS, Pen-Strep (0.1%) and 2-mercaptoethanol (0.014 v/v) and kept in a 37° C CO2 incubator in filter-cap flasks.   Genomic DNA was isolated with aDNeasy Blood & Tissue Kit (Qiagen, Hilden) according to the manufacturer's protocol. The RNAi cassettes remaining in the PHT39-selected clones were amplified from genomic DNA using the LIB2 primer pair (PMID: 25502887). PCR products were purified (PCR purification kit, Qiagen) and subjected to PCR-free library preparation.
description
not collected
geographic location (country and/or sea)
not applicable
isolate
Trypanosoma brucei TREU927
organism
Trypanosoma brucei brucei TREU927
organism
whole organism
organism part
Trypanosoma brucei brucei TREU927
scientific_name
PH39
title