Molecular Biology

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    Protocols in Current Issue
    Imaging Higher-order Chromatin Structures in Single Cells Using Stochastic Optical Reconstruction Microscopy
    Authors:  Jianquan Xu and Yang Liu, date: 02/05/2019, view: 2007, Q&A: 0
    [Abstract] Higher-order chromatin organization shaped by epigenetic modifications influence the chromatin environment and subsequently regulate gene expression. Direct visualization of the higher-order chromatin structure at their epigenomic states is of great importance for understanding chromatin compaction and its subsequent effect on gene expression and ...
    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: 3106, 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 ...
    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: 6243, 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 ...
    In vitro Engineered DNA-binding Molecule-mediated Chromatin Immunoprecipitation (in vitro enChIP) Using CRISPR Ribonucleoproteins in Combination with Next-generation Sequencing (in vitro enChIP-Seq) for the Identification of Chromosomal Interactions
    Authors:  Toshitsugu Fujita and Hodaka Fujii, date: 11/20/2017, view: 4988, Q&A: 0
    [Abstract] We have developed locus-specific chromatin immunoprecipitation (locus-specific ChIP) technologies consisting of insertional ChIP (iChIP) and engineered DNA-binding molecule-mediated ChIP (enChIP). Locus-specific ChIP is a method to isolate a genomic region of interest from cells while it also identifies what binds to this region using mass ...
    Nucleosome Positioning Assay
    Authors:  Zhongliang Zhao and Holger Bierhoff, date: 05/20/2017, view: 6574, Q&A: 0
    [Abstract] The basic unit of chromatin is the nucleosome, a histone octamer with 147 base pairs of DNA wrapped around it. Positions of nucleosomes relative to each other and to DNA elements have a strong impact on chromatin structure and gene activity and are tightly regulated at multiple levels, i.e., DNA sequence, transcription factor binding, ...
    Preparation of Mitotic and Meiotic Metaphase Chromosomes from Young Leaves and Flower Buds of Coccinia grandis
    [Abstract] Somatic chromosomes are usually studied from the root tip cells of the plants for cytological investigations. In dioecious plant species like Coccinia grandis, it is very difficult to get meristematic root tip cells from the mature plants of the respective sex forms. In this report, young leaves of the respective sexual phenotypes were ...
    DNA Slot Blot Repair Assay
    Authors:  Jeong-Min Park and Tae-Hong Kang, date: 04/20/2015, view: 13500, Q&A: 0
    [Abstract] Ultraviolet (UV) irradiation induces helix distorting photolesions such as cyclobutane pyrimidine dimers (CPD) and pyrimidine-pyrimidone (6-4) photoproducts (6-4PP) which threaten genomic integrity if unrepaired. In mammals, nucleotide excision repair (NER) is the only pathway that removes UV-induced DNA damages. Here we describe DNA slot blot ...
    Rice Meiotic Chromosome Spread Preparation of Pollen Mother Cells
    Authors:  Xingwang Li and Changyin Wu, date: 07/20/2014, view: 7062, Q&A: 0
    [Abstract] In this protocol, we describe a simple and efficient method for meiotic chromosome spread preparation in rice pollen mother cells. Meiotic chromosome preparation by spreading itself is an important technique for plant cytogenetics (Higgins et al., 2004; Chelysheva et al., 2012; Wang et al., 2009); furthermore, it is a ...
    Protocol for the Preparation of Arabidopsis Meiotic Chromosome Spreads and Fluorescent in situ Hybridization
    Authors:  Pablo Bolaños-Villegas, Xiaohui Yang, Christopher A. Makaroff and Guang-Yuh Jauh, date: 04/20/2014, view: 10872, Q&A: 0
    [Abstract] This protocol is a more detailed version of previous protocols (Yang et al., 2011; Bolaños-Villegas et al., 2013) developed for the examination of meiotic chromosome spreads. Meiotic chromosome spreads are useful to determine the presence of defects in chromosome pairing and segregation. The protocol also describes how to perform ...
    Micrococcal Nuclease (MNase) Assay of Arabidopsis thaliana Nuclei
    Authors:  Laia Armengot and Jordi Moreno-Romero, date: 04/05/2013, view: 16051, Q&A: 2
    [Abstract] Micrococcal nuclease (MNase) is able to produce double-strand breaks within nucleosome linker regions. The efficiency of MNase digestion depends on the degree of chromatin compaction, being more easily digested the regions of less compacted chromatin. The MNase protocol described here can be used to asses changes in the chromatin structure of ...



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