Ontology highlight
ABSTRACT:
PROVIDER: PRJEB14111 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| ERR1418313_1.fastq.gz | Fastqsanger.gz | |||
| ERR1418313_2.fastq.gz | Fastqsanger.gz | |||
| ERR1418314_1.fastq.gz | Fastqsanger.gz | |||
| ERR1418314_2.fastq.gz | Fastqsanger.gz | |||
| ERR1418315_1.fastq.gz | Fastqsanger.gz |
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Project description:We describe XmaI-RRBS method for rapid and affordable genome-wide DNA methylation analysis, with library preparation taking only four days and sequencing possible within four hours. Small sizes of the XmaI-RRBS libraries allow their multiplexing and sequencing on the benchtop high-throughput machines. Described here is the first RRBS protocol validated for the Ion Torrent Personal Genome Machine. DNA from MCF7 cell line and 6 normal breast samples (total 7 samples) were subjected to reduced representation bisulfite sequencing analysis (XmaI-RRBS) by using Ion Torrent platform.
2015-11-01 | E-GEOD-74126 | biostudies-arrayexpress
Project description:We aimed at obtaining a reference transcriptome for the European seabass. We characterized by Illumina paired-ends RNA-seq the D. labrax transcriptome for two targeted organs, using a diversity of conditions, animal stages and genotypes to warranty the widest variety of reconstructed transcripts. The brain (including pineal gland and pituitary) and liver were chosen as they are complementary and at a crossroads of many key neuroendocrine, metabolic and behavioral regulations. Dicentrarchus labrax_2 targeted organs (liver and brain) with two technical replicates each (using two different fragment sizes) half a GAIIX run total. We used two flow cells per organ and each was corresponding to a different fragment size. The expected sizes were 350 and 500bp.
2014-02-01 | E-MTAB-1867 | biostudies-arrayexpress
Project description:Purpose: The goal of this study is to use chromatin accessibility data to study the information patterns of chromatin accessibility that encode TF-chromatin interactions. Methods: We generated chromatin accessibility data from the GM12878 lymphoblastoid cell line using a modified protocol where the ATAC-seq fragments are sonicated during the library preparation. The goal of the sonication step is to disrupt the observed fragment size distribution, therefore masking local TF-chromatin interactions encoded in the larger fragment sizes (e.g. nucleosome phasing) without affecting the levels of chromatin accessibility.
2020-12-21 | GSE135074 | GEO
Project description:We describe XmaI-RRBS method for rapid and affordable genome-wide DNA methylation analysis, with library preparation taking only four days and sequencing possible within four hours. Small sizes of the XmaI-RRBS libraries allow their multiplexing and sequencing on the benchtop high-throughput machines. Described here is the first RRBS protocol validated for the Ion Torrent Personal Genome Machine.
2015-11-01 | GSE74126 | GEO
Project description:Purpose: to compare different Methyl Binding Domain (MBD) based kits for DNA-methylation sequencing using Reduced Representation Bisulfite Sequencing (RRBS) data for validation, and to determine whether data quality can also be derived from inherent sequence data characteristics MBD-seq using 5 different kits (MethylCap, MethylCollector, MethylCollector Ultra, MethylMiner, MethylMagnet) was applied on 3 commonly used cell lines (DU145, HCT15, PC3), for which also RRBS data were generated.
2013-01-23 | E-GEOD-42790 | biostudies-arrayexpress
Project description:DNA methylation is an epigenetic mechanism that plays roles in gene regulation and expression. This study aims to profile DNA methylation at genome-wide scale and single base resolution using reduced representation bisulfite sequencing (RRBS) of the muscle of pigs differing in muscle metabolism and genetic background. Pietrain homozygous- positive (PiPP) and negative (PiNN) for the ryanodine receptor mutation and malignant hyperthermia-susceptible (MHS), Duroc and F2 crossbred Duroc-Pietrain homozygous-negative (DuPi) pigs were investigated to identify differentially methylated CpG sites/ regions and to integrate the results with gene expression data.
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Project description:This data was generated by ENCODE. If you have questions about the data, contact the submitting laboratory directly (Florencia Pauli mailto:fpauli@hudsonalpha.org). If you have questions about the Genome Browser track associated with this data, contact ENCODE (mailto:genome@soe.ucsc.edu). This track is produced as part of the ENCODE project. The track reports the percentage of DNA molecules that exhibit cytosine methylation at specific CpG dinucleotides. In general, DNA methylation within a gene's promoter is associated with gene silencing, and DNA methylation within the exons and introns of a gene is associated with gene expression. Proper regulation of DNA methylation is essential during development and aberrant DNA methylation is a hallmark of cancer. DNA methylation status is assayed at more than 500,000 CpG dinucleotides in the genome using Reduced Representation Bisulfite Sequencing (RRBS). Genomic DNA is digested with the methyl-insensitive restriction enzyme MspI, small genomic DNA fragments are purified by gel electrophoresis, and then used to construct an Illumina sequencing library. The library fragments are treated with sodium bisulfite and amplified by PCR to convert every unmethylated cytosine to a thymidine while leaving methylated cytosines intact. The sequenced fragments are aligned to a customized reference genome sequence and for each assayed CpG we report the number of sequencing reads covering that CpG and the percentage of those reads that are methylated. For data usage terms and conditions, please refer to http://www.genome.gov/27528022 and http://www.genome.gov/Pages/Research/ENCODE/ENCODEDataReleasePolicyFinal2008.pdf DNA methylation at CpG sites was assayed with a modified version of Reduced Representation Bisulfite Sequencing (RRBS; Meissner et al., 2008). RRBS was performed on cell lines grown by many ENCODE production groups. The production group that grew the cells and isolated genomic DNA is indicated in the "obtainedBy" field of the metadata. When a cell type was provided by more than one lab, the data for the cells from only one lab are displayed in the table above. However, the data for every cell type from every lab is available from the Downloads page. RRBS was carried out by the Myers production group at the HudsonAlpha Institute for Biotechnology. Isolation of genomic DNA Genomic DNA is isolated from biological replicates of each cell line using the QIAGEN DNeasy Blood & Tissue Kit according to the instructions provided by the manufacturer. DNA concentrations for each genomic DNA preparation are determined using fluorescent DNA binding dye and a fluorometer (Invitrogen Quant-iT dsDNA High Sensitivity Kit and Qubit Fluorometer). Typically, 1 µg of DNA is used to make an RRBS library; however, we have also had success in making libraries with 200 ng genomic DNA from rare or precious samples. RRBS library construction and sequencing RRBS library construction starts with MspI digestion of genomic DNA , which cuts at every CCGG regardless of methylation status. Klenow exo- DNA Polymerase is then used to fill in the recessed end of the genomic DNA and add an adenosine as a 3prime overhang. Next, a methylated version of the Illumina paired-end adapters is ligated onto the DNA. Adapter ligated genomic DNA fragments between 105 and 185 basepairs are selected using agarose gel electrophoresis and Qiagen Qiaquick Gel Extraction Kit. The selected adapter-ligated fragments are treated with sodium bisulfite using the Zymo Research EZ DNA Methylation Gold Kit, which converts unmethylated cytosines to uracils and leaves methylated cytosines unchanged. Bisulfite treated DNA is amplified in a final PCR reaction which has been optimized to uniformly amplify diverse fragment sizes and sequence contexts in the same reaction. During this final PCR reaction uracils are copied as thymines resulting in a thymine in the PCR products wherever an unmethylated cytosine existed in the genomic DNA. The sample is now ready for sequencing on the Illumina sequencing platform. These libraries were sequenced with an Illumina Genome Analyzer IIx according to the manufacturer's recommendations. Data analysis To analyze the sequence data, a reference genome is created that contains only the 36 base pairs adjacent to every MspI site and every C in those sequences is changed to T. A converted sequence read file is then created by changing each C in the original sequence reads to a T. The converted sequence reads are aligned to the converted reference genome, and only reads that map uniquely to the reference genome are kept. Once reads are aligned the percent methylation is calculated for each CpG using the original sequence reads. The percent methylation and number of reads is reported for each CpG.
2011-03-01 | E-GEOD-27584 | biostudies-arrayexpress
Project description:Boar taint (BT) is an offensive flavor observed in non-castrated male pigs that reduces the carcass price. Surgical castration effectively avoids the taint but is associated with animal welfare concerns. The functional annotation of farm animal genomes (FAANG) for understanding the biology of complex traits and diseases can be used in the selection of breeding animals to achieve favorable phenotypic outcomes. The characterization of pig epigenomes/methylation changes between animals with high and low BT and genome-wide epigenetic markers that can predict BT are lacking. Reduced representation bisulfite sequencing (RRBS) of DNA methylation patterns based on next generation sequencing (NGS) is an efficient technology to identify candidate epigenetic biomarkers associated with BT. Three different BT levels were analyzed using RRBS data to calculate the methylation levels of cytosine. The co-analysis of differentially methylated cytosines (DMCs) identified by this study and differentially expressed (DE) genes identified by previous study found 32 significant co-located genes. The joint analysis of GO terms and pathways revealed that methylation and gene expression of seven candidate genes were associated with BT; in particular, FASN plays a key role in fatty acid biosynthesis, and PEMT might be involved in estrogen regulation and the development of BT. This study is the first to report the genome-wide DNA methylation profiles of BT in pigs using NGS and summarize candidate genes associated with epigenetic markers of BT, which could contribute to the understanding of functional biology of BT traits and selective breeding of pigs against BT based on epigenetic biomarkers.
2019-10-01 | GSE129385 | GEO
Project description:For data-independent acquisition by means of sequential window acquisition of all theoretical fragment ion spectra (SWATH), a reference library of data-dependent acquisition (DDA) runs is typically used to correlate the quantitative data from the fragment ion spectra with peptide identifications. The quality and coverage of such a reference library is therefore essential when processing SWATH data. In general, library sizes can be increased by reducing the impact of DDA precursor selection with replicate runs or fractionation. However, these strategies can affect the match between the library and SWATH measurement, and thus larger library sizes do not necessarily correspond to improved SWATH quantification. Here, three fractionation strategies to increase local library size were compared to standard library building using replicate DDA injection: protein SDS-PAGE fractionation, peptide high-pH RP-HPLC fractionation and MS-acquisition gas phase fractionation. The impact of these libraries on SWATH performance was evaluated in terms of the number of extracted peptides and proteins, the match quality of the peptides and the extraction reproducibility of the transitions. These analyses were conducted using the hydrophilic proteome of differentiating human embryonic stem cells.
2017-07-04 | PXD006190 | Pride