| FRANEK, Michal, Agata Magdalena KILAR, Petr FOJTÍK, Marie OLSINOVA, Ales BENDA, Vladimír ROTREKL, Martina DVOŘÁČKOVÁ and Jiří FAJKUS. Super-resolution microscopy of chromatin fibers and quantitative DNA methylation analysis of DNA fiber preparations. Journal of Cell Science. Cambridge: Company of Biologists Ltd., 2021, vol. 134, No 15, p. 1-14. ISSN 0021-9533. Available from: https://dx.doi.org/10.1242/jcs.258374. |
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@article{1836817,
author = {Franek, Michal and Kilar, Agata Magdalena and Fojtík, Petr and Olsinova, Marie and Benda, Ales and Rotrekl, Vladimír and Dvořáčková, Martina and Fajkus, Jiří},
article_location = {Cambridge},
article_number = {15},
doi = {http://dx.doi.org/10.1242/jcs.258374},
keywords = {DNA; Fiber; Chromatin; Methylation; Microscopy},
language = {eng},
issn = {0021-9533},
journal = {Journal of Cell Science},
title = {Super-resolution microscopy of chromatin fibers and quantitative DNA methylation analysis of DNA fiber preparations},
url = {https://journals.biologists.com/jcs/article/134/15/jcs258374/271206/Super-resolution-microscopy-of-chromatin-fibers},
volume = {134},
year = {2021}
}
TY - JOUR
ID - 1836817
AU - Franek, Michal - Kilar, Agata Magdalena - Fojtík, Petr - Olsinova, Marie - Benda, Ales - Rotrekl, Vladimír - Dvořáčková, Martina - Fajkus, Jiří
PY - 2021
TI - Super-resolution microscopy of chromatin fibers and quantitative DNA methylation analysis of DNA fiber preparations
JF - Journal of Cell Science
VL - 134
IS - 15
SP - 1-14
EP - 1-14
PB - Company of Biologists Ltd.
SN - 00219533
KW - DNA
KW - Fiber
KW - Chromatin
KW - Methylation
KW - Microscopy
UR - https://journals.biologists.com/jcs/article/134/15/jcs258374/271206/Super-resolution-microscopy-of-chromatin-fibers
N2 - Analysis of histone variants and epigenetic marks is dominated by genome-wide approaches in the form of chromatin immunoprecipitation-sequencing (ChIP-seq) and related methods. Although uncontested in their value for single-copy genes, mapping the chromatin of DNA repeats is problematic for biochemical techniques that involve averaging of cell populations or analysis of clusters of tandem repeats in a single-cell analysis. Extending chromatin and DNA fibers allows us to study the epigenetics of individual repeats in their specific chromosomal context, and thus constitutes an important tool for gaining a complete understanding of the epigenetic organization of genomes. We report that using an optimized fiber extension protocol is essential in order to obtain more reproducible data and to minimize the clustering of fibers. We also demonstrate that the use of super-resolution microscopy is important for reliable evaluation of the distribution of histone modifications on individual fibers. Furthermore, we introduce a custom script for the analysis of methylation levels on DNA fibers and apply it to map the methylation of telomeres, ribosomal genes and centromeres.
ER -
FRANEK, Michal, Agata Magdalena KILAR, Petr FOJTÍK, Marie OLSINOVA, Ales BENDA, Vladimír ROTREKL, Martina DVOŘÁČKOVÁ and Jiří FAJKUS. Super-resolution microscopy of chromatin fibers and quantitative DNA methylation analysis of DNA fiber preparations. \textit{Journal of Cell Science}. Cambridge: Company of Biologists Ltd., 2021, vol.~134, No~15, p.~1-14. ISSN~0021-9533. Available from: https://dx.doi.org/10.1242/jcs.258374.
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