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Quantitative ChIP-seq by Adding Spike-in from Another Species

Authors: Kongyan Niu, Rui Liu and Nan Liu, date: 08/20/2018, view: 4412, Q&A: 0

[Abstract] Chromatin immunoprecipitation followed by sequencing (ChIP-seq) is a routine procedure in the lab; however, epigenome-wide quantitative comparison among independent ChIP-seq experiments remains a challenge. Here, we contribute an experimental protocol combined with a computational workflow allowing quantitative and comparative assessment of ...

Tethered Chromosome Conformation Capture Sequencing in Triticeae: A Valuable Tool for Genome Assembly

Authors: Axel Himmelbach, Ines Walde, Martin Mascher and Nils Stein, date: 08/05/2018, view: 3081, Q&A: 0

[Abstract] Chromosome conformation capture sequencing (Hi-C) is a powerful method to comprehensively interrogate the three-dimensional positioning of chromatin in the nucleus. The development of Hi-C can be traced back to successive increases in the resolution and throughput of chromosome conformation capture (3C) (Dekker et al., 2002). The basic ...

Quantifying Symmetrically Methylated H4R3 on the Kaposi’s Sarcoma-associated Herpesvirus (KSHV) Genome by ChIP-Seq

Authors: Roxanne C. Strahan, Kayla S. Hiura and Subhash C. Verma, date: 03/20/2018, view: 3182, Q&A: 0

[Abstract] Post-translational modifications to histone tails contribute to the three-dimensional structure of chromatin and play an important role in determining the relative expression of nearby genes. One such modification is symmetric di-methylation of arginine residues, which may exhibit different effects on gene expression including blocking the binding ...

Reduced Representation Bisulfite Sequencing in Maize

Authors: Fei-Man Hsu, Chung-Ju Rachel Wang and Pao-Yang Chen, date: 03/20/2018, view: 3459, Q&A: 0

[Abstract] DNA methylation is an epigenetic modification that regulates plant development (Law and Jacobsen, 2010). Whole genome bisulfite sequencing (WGBS) is a state-of-the-art method for profiling genome-wide methylation patterns with single-base resolution (Cokus et al., 2008). However, for an organism with a large genome, e.g., the 2.1 ...

Coupling Exonuclease Digestion with Selective Chemical Labeling for Base-resolution Mapping of 5-Hydroxymethylcytosine in Genomic DNA

Authors: Aurélien A. Sérandour, Stéphane Avner and Gilles Salbert, date: 03/05/2018, view: 3272, Q&A: 0

[Abstract] This protocol is designed to obtain base-resolution information on the level of 5-hydroxymethylcytosine (5hmC) in CpGs without the need for bisulfite modification. It relies on (i) the capture of hydroxymethylated sequences by a procedure known as ‘selective chemical labeling’ (see Szulwach et al., 2012) and (ii) the digestion of the ...

Low-input Capture-C: A Chromosome Conformation Capture Assay to Analyze Chromatin Architecture in Small Numbers of Cells

Authors: A. Marieke Oudelaar, Damien J. Downes, James O.J. Davies and Jim R. Hughes, date: 12/05/2017, view: 6216, Q&A: 0

[Abstract] Chromosome conformation capture (3C) techniques are crucial to understanding tissue-specific regulation of gene expression, but current methods generally require large numbers of cells. This protocol describes two new low-input Capture-C approaches that can generate high-quality 3C interaction profiles from 10,000-20,000 cells, depending on the ...

Bioinformatic Analysis for Profiling Drug-induced Chromatin Modification Landscapes in Mouse Brain Using ChlP-seq Data

Authors: Yong-Hwee Eddie Loh, Jian Feng, Eric Nestler and Li Shen, date: 02/05/2017, view: 6072, Q&A: 0

[Abstract] Chromatin immunoprecipitation followed by massively parallel sequencing (ChIP-seq) is a powerful technology to profile genome-wide chromatin modification patterns and is increasingly being used to study the molecular mechanisms of brain diseases such as drug addiction. This protocol discusses the typical procedures involved in ChIP-seq data ...

Whole Genome Bisulfite Sequencing and DNA Methylation Analysis from Plant Tissue

Authors: Daniela Pignatta, George W. Bell and Mary Gehring, date: 02/20/2015, view: 13939, Q&A: 0

[Abstract] This protocol describes whole genome bisulfite-sequencing library preparation from plant tissue and subsequent data analysis. Allele-specific methylation analysis and genome-wide identification of differentially methylated regions are additional features of the analysis procedure.

Library Construction for Genome-wide Bisulfite Sequencing in Plants

Authors: David Moraga Amador, Chenggang Wang, Kevin H. Holland and Zhonglin Mou, date: 12/20/2013, view: 10219, Q&A: 0

[Abstract] DNA methylation is the most studied epigenetic modification, which involves the addition of a methyl group to the carbon-5 position of cytosine residues in DNA. DNA methylation is important for the regulation of gene expression. Bisulfite sequencing is the gold standard technique for determining genome-wide DNA methylation profiles in eukaryotes. ...

Quantitative Methylation Specific PCR (qMSP)

Authors: Triantafillos Liloglou and Georgios Nikolaidis, date: 08/20/2013, view: 22079, Q&A: 0

[Abstract] Detection of low copies of methylated DNA targets in clinical specimens is challenging. The quantitative Methylation-Specific PCR (qMSP) assays were designed to specifically amplify bisulphite-converted methylated DNA target sequences in the presence of an excess of unmethylated counterpart sequences. These qMSP assays are real-time PCR assays ...

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