Systems Biology


    Protocols in Current Issue
    A Molecular Cloning and Sanger Sequencing-based Protocol for Detecting Site-specific DNA Methylation
    Authors:  Wei Guo, Anthony Cannon and Damon Lisch, date: 05/05/2022, view: 519, Q&A: 0

    DNA methylation is a conserved chemical modification, by which methyl groups are added to the cytosine of DNA molecules. Methylation can influence gene expression without changing the sequence of a particular gene. This epigenetic effect is an intriguing phenomenon that has puzzled biologists for years. By probing the temporal and spatial patterns

    Whole-genome Methylation Analysis of APOBEC Enzyme-converted DNA (~5 kb) by Nanopore Sequencing

    In recent years, DNA methylation research has been accelerated by the advent of nanopore sequencers. However, read length has been limited by the constraints of base conversion using the bisulfite method, making analysis of chromatin content difficult. The read length of the previous method combining bisulfite conversion and long-read sequencing

    Histone modification ChIP-seq on Arabidopsis thaliana Plantlets
    Authors:  Juan S. Ramirez-Prado, David Latrasse and Moussa Benhamed, date: 11/05/2021, view: 1700, Q&A: 1

    Characterizing the molecular mechanisms regulating gene expression is crucial for understanding the regulatory processes underlying physiological responses to environmental and developmental signals in eukaryotes. The covalent modification of histones contributes to the compaction levels of chromatin, as well as the recruitment of the

    Assessing the in vitro Binding Specificity of Histone Modification Reader Proteins Using Histone Peptide Arrays
    Authors:  Mark W. Soo and Arneet L. Saltzman, date: 09/20/2021, view: 1147, Q&A: 0

    In the field of chromatin biology, a major goal of understanding the roles of histone post-translational modifications is to identify the proteins and domains that recognize these modifications. Synthetic histone peptides containing one or more modifications are a key tool to probe these interactions in pull-down assays with recombinant proteins

    Isolation of Nuclei from Mouse Dorsal Root Ganglia for Single-nucleus Genomics
    Authors:  Lite Yang, Ivan Tochitsky, Clifford J. Woolf and William Renthal, date: 08/05/2021, view: 1895, Q&A: 0

    Primary somatosensory neurons, whose cell bodies reside in the dorsal root ganglion (DRG) and trigeminal ganglion, are specialized to transmit sensory information from the periphery to the central nervous system. Our molecular understanding of peripheral sensory neurons has been limited by both their heterogeneity and low abundance compared with

    Detecting Differentially Methylated Promoters in Genes Related to Disease Phenotypes Using R
    Authors:  Jordi Martorell Marugán and Pedro Carmona-Sáez, date: 06/05/2021, view: 2010, Q&A: 0

    DNA methylation in gene promoters plays a major role in gene expression regulation, and alterations in methylation patterns have been associated with several diseases. In this context, different software suites and statistical methods have been proposed to analyze differentially methylated positions and regions. Among them, the novel statistical

    Simplified Epigenome Profiling Using Antibody-tethered Tagmentation
    Authors:  Steven Henikoff, Jorja G. Henikoff and Kami Ahmad, date: 06/05/2021, view: 3212, Q&A: 0

    We previously introduced Cleavage Under Targets & Tagmentation (CUT&Tag), an epigenomic profiling method in which antibody tethering of the Tn5 transposase to a chromatin epitope of interest maps specific chromatin features in small samples and single cells. With CUT&Tag, intact cells or nuclei are permeabilized, followed by successive

    Quantitative ChIP-seq by Adding Spike-in from Another Species
    Authors:  Kongyan Niu, Rui Liu and Nan Liu, date: 08/20/2018, view: 18779, 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: 7208, 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: 6116, 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 ...

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