دورية أكاديمية
أعرض في EDS

Formamide denaturation of double-stranded DNA for fluorescence in situ hybridization (FISH) distorts nanoscale chromatin structure.

التفاصيل البيبلوغرافية
العنوان: Formamide denaturation of double-stranded DNA for fluorescence in situ hybridization (FISH) distorts nanoscale chromatin structure.
المؤلفون: Anne R Shim, Jane Frederick, Emily M Pujadas, Tiffany Kuo, I Chae Ye, Joshua A Pritchard, Cody L Dunton, Paola Carrillo Gonzalez, Nicolas Acosta, Surbhi Jain, Nicholas M Anthony, Luay M Almassalha, Igal Szleifer, Vadim Backman
المصدر: PLoS ONE, Vol 19, Iss 5, p e0301000 (2024)
بيانات النشر: Public Library of Science (PLoS), 2024.
سنة النشر: 2024
المجموعة: LCC:Medicine
LCC:Science
مصطلحات موضوعية: Medicine, Science
الوصف: As imaging techniques rapidly evolve to probe nanoscale genome organization at higher resolution, it is critical to consider how the reagents and procedures involved in sample preparation affect chromatin at the relevant length scales. Here, we investigate the effects of fluorescent labeling of DNA sequences within chromatin using the gold standard technique of three-dimensional fluorescence in situ hybridization (3D FISH). The chemical reagents involved in the 3D FISH protocol, specifically formamide, cause significant alterations to the sub-200 nm (sub-Mbp) chromatin structure. Alternatively, two labeling methods that do not rely on formamide denaturation, resolution after single-strand exonuclease resection (RASER)-FISH and clustered regularly interspaced short palindromic repeats (CRISPR)-Sirius, had minimal impact on the three-dimensional organization of chromatin. We present a polymer physics-based analysis of these protocols with guidelines for their interpretation when assessing chromatin structure using currently available techniques.
نوع الوثيقة: article
وصف الملف: electronic resource
اللغة: English
تدمد: 1932-6203
Relation: https://doaj.org/toc/1932-6203
DOI: 10.1371/journal.pone.0301000
URL الوصول: https://doaj.org/article/89f2c803491e4e51a3af9fa3dee2c896
رقم الأكسشن: edsdoj.89f2c803491e4e51a3af9fa3dee2c896
قاعدة البيانات: Directory of Open Access Journals
الوصف
تدمد:19326203
DOI:10.1371/journal.pone.0301000