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
ERS18345197
SRA accession
E-MTAB-13843:NT-3_1
Submitter Id
ArrayExpress
broker name
not collected
collection date
Protocols: T. brucei BSF cells in logarithmic growth phase were treated with 400 nM AN3057 (3 × EC50 determined after 24h) and placed in an ice-water mix for 5 min after 20 min and 60 min drug exposure. 6.5 x 10e7 cells were harvested by centrifugation (1000*g, 4°C) together with nontreated samples prepared in parallel. All samples were in 2 biological replicates.  Bloodstream T. brucei (Lister 427) were propagated in T-25 vented cap flasks (Corning Inc., Lowell, MA) at 37˚C and 5% CO2 with humidity in HMI-9 medium and maintained at a cell density less than 10e6 cell/ml (Hirumi and Hirumi, 1989). Media was supplemented with 10% foetal bovine serum (v/v) and 1% antibiotic solution (v/v) (Sigma Aldrich).  RNA was extracted with the RNeasy RNA extraction kit (Qiagen) following the manufacturer’s protocol. Total RNA yields were 12-16 g with a RNA quality number (RNQ) > 6 RNA samples were stored at −80 °C and sent to BGI for library preparation and sequencing. After oligo dT enrichment, mRNA was fragmented and subjected to first strand cDNA synthesis using random primers.Second strand cDNA was synthesized with dUTP instead of dTTP. The synthesized cDNA was subjected to end-repair and 3' adenylation before adaptor ligation. Upon digestion of the U-labeled second-strand template with uracil-DNA-glycosylase the library was PCR amplified.
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
NT-3_1
title