Please use this identifier to cite or link to this item: doi:10.22028/D291-29412
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Title: Hairpin Bisulfite Sequencing: Synchronous Methylation Analysis on Complementary DNA Strands of Individual Chromosomes
Author(s): Giehr, Peter Pascal
Walter, Jörn Erik
Language: English
Title: Methods in molecular biology
Volume: 1708
Startpage: 573
Endpage: 586
Publisher/Platform: Springer
Year of Publication: 2017
Publikation type: Journal Article
Abstract: The accurate and quantitative detection of 5-methylcytosine is of great importance in the field of epigenetics. The method of choice is usually bisulfite sequencing because of the high resolution and the possibility to combine it with next generation sequencing. Nevertheless, also this method has its limitations. Following the bisulfite treatment DNA strands are no longer complementary such that in a subsequent PCR amplification the DNA methylation patterns information of only one of the two DNA strand is preserved. Several years ago Hairpin Bisulfite sequencing was developed as a method to obtain the pattern information on complementary DNA strands. The method requires fragmentation (usually by enzymatic cleavage) of genomic DNA followed by a covalent linking of both DNA strands through ligation of a short DNA hairpin oligonucleotide to both strands. The ligated covalently linked dsDNA products are then subjected to a conventional bisulfite treatment during which all unmodified cytosines are converted to uracils. During the treatment the DNA is denatured forming noncomplementary ssDNA circles. These circles serve as a template for a locus specific PCR to amplify chromosomal patterns of the region of interest. As a result one ends up with a linearized product, which contains the methylation information of both complementary DNA strands.
DOI of the first publication: 10.1007/978-1-4939-7481-8_29
URL of the first publication:
Link to this record: hdl:20.500.11880/27880
ISSN: 1064-3745
Date of registration: 24-Sep-2019
Faculty: NT - Naturwissenschaftlich- Technische Fakultät
Department: NT - Biowissenschaften
Professorship: NT - Prof. Dr. Jörn Walter
Collections:SciDok - Der Wissenschaftsserver der Universität des Saarlandes

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