Project description:The mRNA expression of antimony resistant strains of Leishmania donovani was compared to the expression of the sensitive Leishmania donovani. The antimony resistant and sensitive Leishmania donovani were grown in complete M199 medium with 10% FCS and Penicillin streptomycin mixture. At stationary phase (5 day culture) cells were harvested in sterile Phosphate buffered saline and used for RNA isolation.
Project description:The aim of the experiment is to detect any gene expression modulation in an antimony resistant L.infantum Sb2000.1 strain compared to an antimony sensitive L.infantum WT strain. We want to identify the modulated genes associated with an antimony resistant phenotype. The Sb2000.1 resistant strain used is about 1000 times more resistant to SbIII than the WT sensitive strain. The full-genome DNA microarrays includes one 70-oligonucleotide probe for each gene of L. infantum. Keywords: drug resistance
Project description:The aim of the experiment is to detect any gene expression modulation in an antimony resistant L.infantum Sb2000.1 strain compared to an antimony sensitive L.infantum WT strain. We want to identify the modulated genes associated with an antimony resistant phenotype. The Sb2000.1 resistant strain used is about 1000 times more resistant to SbIII than the WT sensitive strain. The full-genome DNA microarrays includes one 70-oligonucleotide probe for each gene of L. infantum. Keywords: drug resistance Two-condition experiment, resistant strain Sb2000.1 vs wild-type. Five biological replicates for each strain, independently grown and harvested. One replicate per array
Project description:Contaminated aquifer (Dusseldorf-Flinger, Germany) templates extracted from 5 sediment depths ranging between 6.4 and 8.4 m below ground and over 3 years of sampling were amplified for amplicon pyrosequencing using the primers Ba27f (5’-aga gtt tga tcm tgg ctc ag-3’) and Ba519r (5’- tat tac cgc ggc kgc tg-3’), extended as amplicon fusion primers with respective primer A or B adapters, key sequence and multiplex identifiers (MID) as recommended by 454/Roche. Amplicons were purified and pooled as specified by the manufacturer. Emulsion PCR (emPCR), purification of DNA-enriched beads and sequencing run were performed following protocols and using a 2nd generation pyrosequencer (454 GS FLX Titanium, Roche) as recommended by the developer. Quality filtering of the pyrosequencing reads was performed using the automatic amplicon pipeline of the GS Run Processor (Roche), with a slight modification concerning the valley filter (vfScanAllFlows false instead of TiOnly) to extract the sequences. Demultiplexed raw reads were furhter trimmed for quality and lenght (>250 bp).