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Universal NicE-Seq: A Simple and Quick Method for Accessible Chromatin Detection in Fixed Cells.
- Source :
-
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2023; Vol. 2611, pp. 39-52. - Publication Year :
- 2023
-
Abstract
- Genome-wide accessible chromatin sequencing and identification has enabled deciphering the epigenetic information encoded in chromatin, revealing accessible promoters, enhancers, nucleosome positioning, transcription factor occupancy, and other chromosomal protein binding. The starting biological materials are often fixed using formaldehyde crosslinking. Here, we describe accessible chromatin library preparation from low numbers of formaldehyde-crosslinked cells using a modified nick translation method, where a nicking enzyme nicks one strand of DNA and DNA polymerase incorporates biotin-conjugated dATP, dCTP, and methyl-dCTP. Once the DNA is labeled, it can be isolated for NGS library preparation. We termed this method as universal NicE-seq (nicking enzyme-assisted sequencing). We also demonstrate a single tube method that enables direct NGS library preparation from low cell numbers without DNA purification. Furthermore, we demonstrated universal NicE-seq on FFPE tissue section sample.<br /> (© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)
Details
- Language :
- English
- ISSN :
- 1940-6029
- Volume :
- 2611
- Database :
- MEDLINE
- Journal :
- Methods in molecular biology (Clifton, N.J.)
- Publication Type :
- Academic Journal
- Accession number :
- 36807062
- Full Text :
- https://doi.org/10.1007/978-1-0716-2899-7_3