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).
Project description:Nitrate-reducing iron(II)-oxidizing (NDFO) bacteria are widespread in the environment contribute to nitrate removal and influence the fate of the greenhouse gases nitrous oxide and carbon dioxide. The autotrophic growth of nitrate-reducing iron(II)-oxidizing bacteria is rarely investigated and poorly understood. The most prominent model system for this type of studies is enrichment culture KS, which originates from a freshwater sediment in Bremen, Germany. A second NDFO culture, culture BP, was obtained with a sample taken in 2015 at the same pond and cultured in a similar way. To gain insights in the metabolism of nitrate reduction coupled to iron(II) oxidation under in the absence of organic carbon and oxygen limited conditions, we performed metagenomic, metatranscriptomic and metaproteomic analyses of culture BP. Raw sequencing data of 16S rRNA amplicon sequencing (V4 region with Illumina and near full-length with PacBio), shotgun metagenomics, metagenome assembly, raw sequencing data of shotgun metatranscriptomes (2 conditions, triplicates) can be found at SRA in https://www.ncbi.nlm.nih.gov/bioproject/PRJNA693457. This dataset contains proteomics data for 2 conditions in triplicates. Samples R23, R24, and R25 are grown in autotrophic conditions, samples R26, R27, and R28 in heterotrophic conditions.
Project description:This study used an emerging analytical technology (cDNA microarrays) to assess the potential effects of PFC exposure on largemouth bass in TCMA lakes. Microarrays simultaneously measure the expression of thousands of genes in various tissues from organisms exposed to different environmental conditions. From this large data set, biomarkers (i.e., genes that are expressed in response to an exposure to known stressors) and bioindicators (e.g., suites of genes that correspond to changes in organism health) can be simultaneously measured to clarify the relationship between contaminant exposure and organism health. Based on current scientific literature, we hypothesized that gene expression patterns would be altered in fish exposed to PFCs (as compared with fish from reference lakes), and that the magnitude of these changes would correspond to the concentrations of PFCs present throughout TCMA lakes. Patterns of gene expression in largemouth bass observed across the TCMA lakes corresponded closely with PFC concentration. Concentrations of PFCs in largemouth bass varied significantly across the sampled lakes, where the lowest concentrations were found in Steiger and Upper Prior Lakes and the highest concentrations were found in Calhoun and Twin Lakes. Patterns of gene expression were most different (relative to controls) in fish with the highest PFC tissue concentrations, where fish from Twin and Calhoun Lakes were observed to have between 5437 and 5936 differentially expressed genes in liver and gonad tissues. Although gene expression patterns demonstrated a high degree of correlation with PFC concentrations, microarray data also suggest there are likely additional factors influencing gene expression patterns in largemouth bass in TCMA lakes.
2014-07-31 | GSE57684 | GEO
Project description:16S and 18S rRNA Amplicon sequencing from Grosses Heiliges Meer, Germany