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DNA methylation at transcription start sites blocks TBP binding and transcription onset by RNApolII [ChIPbisSeq]

ABSTRACT: We report the application of ChIP bisulfite sequencing (ChIPbisSeq) to establish the methylation state of DNA bound to RNApol2 phosphorylated in Ser5 (RNAPol2Ser5) in the mouse cortex. We first profiled the RNAPol2Ser5 binding using regular ChIPSeq from 45 million raw read pairs (31 million unique pairs were aligned to the genome). Then we used bisulfite converted DNA immunoprecipitated with an antibody against RNAPol2Ser5 (87 million raw read pairs, 38 million unique read pairs aligned to the genome) to map RNAPol2Ser5 sites and to establish the methylation state at these sites.

ORGANISM(S): Mus musculus

PROVIDER: GSE60657 | GEO | 2015/09/01

SECONDARY ACCESSION(S): PRJNA259205

REPOSITORIES: GEO

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DNA methylation at transcription start sites blocks TBP binding and transcription onset by RNApolII [ChIPbisSeq]

Project description:We report the application of ChIP bisulfite sequencing (ChIPbisSeq) to establish the methylation state of DNA bound to RNApol2 phosphorylated in Ser5 (RNAPol2Ser5) in the mouse cortex. We first profiled the RNAPol2Ser5 binding using regular ChIPSeq from 45 million raw read pairs (31 million unique pairs were aligned to the genome). Then we used bisulfite converted DNA immunoprecipitated with an antibody against RNAPol2Ser5 (87 million raw read pairs, 38 million unique read pairs aligned to the genome) to map RNAPol2Ser5 sites and to establish the methylation state at these sites. Examination of methylation state at RNAPol2Ser5 binding sites in the mouse cortex.

2015-09-01 | E-GEOD-60657 | biostudies-arrayexpress

Transcriptome of normal, paracancerouse and cancerous bladder tissues based on RNA-Seq method

Project description:In order to support our research of bladder cancer in human genome, we conducted massively parallel pyrosequencing of mRNAs (RNA-Seq) using normal, paracancerouse and cancerous human bladder tissues. We obtained a total of 30.0 million read pairs from normal, 33.1 million read pairs from paracancerous and 36.5 million read pairs from cancerous. The RNA-Seq data derived from the sample illustrated the differencially expression genes among normal, paracancerous and cancerous bladder tissues of human.

2013-01-01 | GSE35178 | GEO

Transcriptome of normal, paracancerouse and cancerous bladder tissues based on RNA-Seq method

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Large conserved domains of low DNA methylation maintained by 5-hydroxymethycytosine and Dnmt3a [Bisulfite-Seq]

Project description:Gains and losses in DNA methylation are prominent genomic features of all mammalian cell types. To gain insight into mechanisms that could promote shifts in DNA methylation patterns and thus contribute to cell fate, including malignant transformation, we performed genome-wide mapping of 5-methylcytosine in purified wild type and Dnmt3a conditional knockout hematopoietic stem cells (HSCs). We generated 1,121 million (control HSCs), and 1,213 million (Dnmt3a knockout HSCs) raw reads; about 81.4% and 88.7%, respectively, were successfully aligned to either strand of the reference genome (mm9), obtainig an average CG coverage of 39.5-fold (wild type) and 47.0-fold (Dnmt3a kockout). Whole genome bisulfite sequencing of secondarily-transplanted wild-type and Dnmt3a conditional knockout hematopoietic stem cells using Illumina HiSeq 2000

2013-11-17 | E-GEOD-49714 | biostudies-arrayexpress

Stallion sperm transcriptome as revealed by microarray analysis and RNA sequencing

Project description:Purpose: In order to understand the functional significance of sperm transcriptome in stallion fertility, the aim of this study was to generate a detailed body of knowledge about the sperm RNA profile that defines a normal fertile stallion. Methods: The 50 bp single-end ABI SOLiD raw reads were directly aligned with the horse reference sequence EcuCab2 using ABI aligner software (NovoalignCS version 1.00.09, novocraft.com) which uses multiple indexes in the reference genome, identifies candidate alignment locations for each primary read, and allows completion of the alignment. Results: Next generation sequencing (NGS) of total RNA from the sperm of two reproductively normal stallions generated about 70 million raw reads and more than 3 Gb of sequence per sample; over half of these aligned with the EcuCab2 reference genome. Altogether, 19,257 sequence tags with average coverage ?1 (normalized number of transcripts) were mapped in the horse genome. Conclusion: The sequence of stallion sperm transcriptome is an important foundation for the discovery of transcripts of known and novel genes, and non-coding RNAs, thus improving the annotation of the horse genome sequence draft and providing markers for evaluating stallion fertility. Reproductively fertile Stallion sperm transcriptome as revealed by RNA sequencing

2013-02-19 | E-GEOD-38725 | biostudies-arrayexpress

Discovery of cis-spliced chimeric RNAs between adjacent genes in human prostate cells

Project description:Total RNA extracted from prostate cancer LNCaP cells transfected with siRNA against CTCF(siCTCF), or negative control siRNA (si-)were processed, and sequenced by two different companies using Illumina Hi-seq 2000 platform to generate RNA sequencing with two output sequences: paired-end 50bp and 101bp in read length. Nearly 100 million and 50 million raw reads were yielded from each sample respectively. We used FastQC to confirm the quality of raw fastq sequencing data, and SOAPfuse software to detect fusion transcripts.

2015-02-09 | GSE63487 | GEO

Cis-SAGe fusion RNAs in transcription splicing factors knocking-down 293T cells

Project description:Total RNA extracted from 15 knocking down treated 293T cells using siRNAs targeting transcription splicing factors and sequenced using Illumina Hiseq PE150 platform to generate RNA sequencing with 150bp in read length. Nearly 50 million raw reads were yielded from each sample respectively. We used FastQC to confirm the quality of raw fastq sequencing data, and Ericscript software to detect fusion transcripts.

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Chimeric RNA profiling demonstrates an association of alveolar rhabdomyosarcoma with specific stages of normal myogenesis

Project description:Total RNA extracted from differentiated mesenchymal stem cells at four time points (T1,T2,T3,T4) and sequenced using Illumina Hi-seq 2000 platform to generate RNA sequencing with 101bp in read length. Nearly 50 million raw reads were yielded from each sample respectively. We used FastQC to confirm the quality of raw fastq sequencing data, and SOAPfuse software to detect fusion transcripts.

2016-02-09 | GSE64032 | GEO

Stallion sperm transcriptome as revealed by microarray analysis and RNA sequencing

Project description:Purpose: In order to understand the functional significance of sperm transcriptome in stallion fertility, the aim of this study was to generate a detailed body of knowledge about the sperm RNA profile that defines a normal fertile stallion. Methods: The 50 bp single-end ABI SOLiD raw reads were directly aligned with the horse reference sequence EcuCab2 using ABI aligner software (NovoalignCS version 1.00.09, novocraft.com) which uses multiple indexes in the reference genome, identifies candidate alignment locations for each primary read, and allows completion of the alignment. Results: Next generation sequencing (NGS) of total RNA from the sperm of two reproductively normal stallions generated about 70 million raw reads and more than 3 Gb of sequence per sample; over half of these aligned with the EcuCab2 reference genome. Altogether, 19,257 sequence tags with average coverage ≥1 (normalized number of transcripts) were mapped in the horse genome. Conclusion: The sequence of stallion sperm transcriptome is an important foundation for the discovery of transcripts of known and novel genes, and non-coding RNAs, thus improving the annotation of the horse genome sequence draft and providing markers for evaluating stallion fertility.

2013-02-19 | GSE38725 | GEO

Discovery of cis-spliced chimeric RNAs between adjacent genes in human prostate cells

2015-02-09 | E-GEOD-63487 | biostudies-arrayexpress

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