Methylome and Hydroxymethylome of iPSC
Ontology highlight
ABSTRACT:
INSTRUMENT(S): Illumina HiSeq 2500
ORGANISM(S): Mus musculus
SUBMITTER:
PROVIDER: E-ERAD-208 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
Similar Datasets
Project description:Mouse B cells were isolated from the spleen and activated ex vivo. DNA was obtained from activated and control B cells with the Qiagen AllPrep kit. 1ug of DNA was sonicated with the Covaris Sonicator (300bp) and the Illumina library was prepared following a standard Illumina protocol with the NEB Next kit. The library was amplified 15x and gel-excised within the range of 200-500bp. This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/
2013-03-25 | E-ERAD-90 | biostudies-arrayexpress
Project description:Mouse Embryonic Stem Cells (ESCs) are grown in culture and seperated based upon heterogeneous expression of key pluripotency factors. DNA was obtained from cells with the Qiagen AllPrep kit. 0.2ug of DNA was sonicated with the Covaris Sonicator (300bp) and the Illumina library was prepared following a standard Illumina protocol with the NEB Next kit. The library was amplified 15x and size-selected within the range of 200-500bp. This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/
2013-08-07 | E-ERAD-178 | biostudies-arrayexpress
Project description:Mouse Epiblast Stem Cells (EpiSCs) were reprogrammed to induced pluripotent stem cells (iPSCs) by overexpression of key pluripotency factors and the TET1 DNA hydroxylase. 0.2ug of extractd DNA was sonicated using the Covaris Sonicator (300bp) and the Illumina library was prepared following a standard Illumina protocol with the NEB Next kit and subjected to bisulfite treament. The library was amplified 12x and fragments smaller than 200 bp were removed by PEG precipitation on magnetic beads. This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/
2014-04-28 | E-ERAD-238 | biostudies-arrayexpress
Project description:Islets were collected and DNA & RNA was extracted using the AllPrep MicroKit. RNA libraries were generated using the ScriptSeq V2 Kit. Libraries were cleaned up with AMPure Beads. BS Libraries were generated using the SigmaImprint Kit. Libraries are barcoded/This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/
2014-10-15 | E-ERAD-302 | biostudies-arrayexpress
Project description:The goal of this study is to identify genomic signatures predicitve of cell-of-origin in acute myeloid leukemia 50K bulk leukemia (GFP+) cells from the spleen of recipient mice were sorted directly into 350μl of RLT buffer (Qiagen) and flash-frozen. Total RNA was isolated according to manufacturer’s protocols (Qiagen) including DNase treatment, and quality was assessed using an Agilent 2100 Bioanalyzer and RNA 6000 Nano kit. Amplified cDNA was sheared to approximately 300bp using a Covaris E220 Focused Ultrasonicator. RNA-seq library preparation used the TruSeq DNA sample prep kit v2 (Illumina). Libraries were sequenced on the Illumina HiSeq 2000 platform.
2016-07-12 | GSE74690 | GEO
Project description:This study is a collaboration with Phil Spence and Jean Langhorne at NIMR, Mill Hill where Phil is studying the differences in aetiology of rodent malaria when mice are infected by mosquito bite versus intraperitoneal injection of blood stage parasites. The aim is to determining transcriptomic differences using RNA-Seq between parasites transmitted by mosquito and those transmitted by injection. This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/Protocol: Mice were bled out at 6 days post-infection, and RNA was extracted from purified parasite populations using Trizol reagent and DNase treated. Poly A+ mRNA was purified from total RNA using oligo dT dyna bead selection and libraries were created using a modified RNA-seq protocol, where RNA was fragmented using Covaris AFA sonication instead of metal ions. The samples were sequenced on an Illumina HiSeq 2000.
2012-11-27 | E-ERAD-95 | biostudies-arrayexpress
Project description:We performed chromatin immunoprecipitation according to standard protocols. Mouse embryonic stem cells were treated with formaldehyde to cross-link proteins to DNA. Cell lysates were sonicated to break up DNA. The histone deubiquitinase Mysm1 was then immunoprecipitated from the sonicated cell lysates. Mysm1-bound (and control antibody-bound) DNA fragments were purified after proteinase K treatment and cross-link reversal. Input DNA was purified directly from the sonicated cell lysates. This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/
2015-11-03 | E-ERAD-398 | biostudies-arrayexpress
Project description:CTCF is a highly conserved and ubiquitously expressed protein involved in several fundamental processes such as fine-tuning gene expression, imprinting, X-chromosome inactivation and 3D chromatin organisation. To understand the impact of differences in the concentration of CTCF abundance on these processes, we exploit a CTCF hemizygous mouse model with a stable reduction in the concentration of this protein. We derived twelve independent primary lines of mouse embryonic fibroblasts (MEFs) from six wildtype and six CTCF-hemizygous mouse E13.5 embryos. MEFs were fixed in DMEM containing 1% fresh formaldehyde and incubated at room temperature for 10 min, quenched with 250mM glycine for 10 min, and washed twice with ice cold PBS, before being flash-frozen at -80°C. Cross-linked cells were lysed and sonicated on a Bioruptor Plus (Diagenode) sonicator to fragment chromatin to an average length of 300bp. 10 ug of the following antibodies were used for immunoprecipitation: CTCF (rabbit polyclonal, Merk Millipore 07-729, lot 2517762); H3K4me3 (mouse monoclonal IgG clone CMA304, Merck Millipore 05-1339, lot 2603814); H3K27ac (rabbit polyclonal IgG, Abcam 4729, lot GR244014-1). Immunoprecipitated DNA or 50 ng of input DNA was used for library preparation using the ThruPLEX DNA-Seq library preparation protocol (Rubicon Genomics, UK). Library fragment size was determined using a 2100 Bioanalyzer (Agilent). Libraries were quantified by qPCR (Kapa Biosystems). Pooled libraries were sequenced on a HiSeq4000 (Illumina) according to manufacturer’s instructions using single-end 50 bp reads. On the same MEF lines we have performed RNAseq and HiC (see related accession numbers).
2018-07-29 | E-MTAB-6261 | biostudies-arrayexpress
Project description:This study focuses on epigenetic reprogramming in the mouse germ line: DNA methylation marks, including those of imprinted genes, are thought to be erased between E11.5 and E13.5 in primordial germ cells (PGCs), which are the direct progenitors of sperm or oocyte. DNA methylation patterns are then re-established during the de novo methylation phase in the male germ line several days later (around E15.5) and in the female germ line after birth during adult life. Epigenetic reprogramming in PGCs is poorly understood mainly because of the technical challenges that arise from very low cell numbers in the embryo. The aim of this study is to create genome-wide maps of DNA methylation patterns of male and female PGCs at crucial time points during epigenetic reprogramming and to investigate the changes in those profiles on a single-gene level. We have already successfully prepared and sequenced the first set of BS-Seq libraries of PGCs with Illumina's GAIIx platform. From this, we gained significant insight into the global DNA methylation levels and methylation patterns over multy-copy-loci such as repeat elements. In order further enhance our analysis and finish this project for publication, it is crucial to increase the genomic coverage of our datasets. This will also allow us to link the BS-Seq data with other MeDIP-Seq and RNA-Seq datasets that have already been created in this study.Protocol: Input DNA was sonicated using a Bioruptor UCD-200 (Diagenode) to a final size distribution of 300bp 1000bp. End-repair and A-tailing were performed with the NEBNext DNA Sample Prep Master Mix Set 1. Illuminas Early Access Methylation Adaptor Oligo Kit was used for the adapter ligation. The adapter-ligated DNA was treated with sodium-bisulfite using the Imprint DNA Modification Kit from Sigma-Aldrich according to the manufacturers instructions for the two-step protocol. After the clean up, the bisulfite-treated DNA was amplified using PfuTurbo Cx Hotstart DNA Polymerase from Agilent with 18 cycles of amplification. The entire PCR reaction was run on a 1.5% agarose TBE gel and size selection was performed for DNA fragments between 200bp 250bp. DNA from the excised gel piece was purified with the Qiagen Gel Extraction Kit.This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/
2013-01-07 | E-ERAD-48 | biostudies-arrayexpress
Project description:Total RNA was extracted from adult male and female zebrafish. Illumina cDNA libraries were prepared from polyA+ RNA and sequenced using Illumina HiSeq paired-end sequencing. This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/ protocol: Total RNA was extracted from zebrafish embryos using Trizol and DNase treated. Libraries were made using the Illumina cDNA protocol from polyA+ RNA.This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/
2013-01-07 | E-ERAD-54 | biostudies-arrayexpress