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Methods Mol Biol. 2018;1708:665-678. doi: 10.1007/978-1-4939-7481-8_34.

Multiplexing for Oxidative Bisulfite Sequencing (oxBS-seq).

Kirschner K1,2,3, Krueger F4, Green AR1,2,3,5, Chandra T6,7,8.

Author information

1
Cambridge Institute for Medical Research, University of Cambridge, Cambridge, CB2 0XY, UK.
2
Department of Haematology, University of Cambridge, Cambridge, UK.
3
Stem Cell Institute, University of Cambridge, Cambridge, UK.
4
Bioinformatics, The Babraham Institute, Cambridge, CB22 3AT, UK.
5
Department of Haematology, Addenbrooke's Hospital, Cambridge, UK.
6
Epigenetics ISP, The Babraham Institute, Cambridge, CB22 3AT, UK. tamir.chandra@igmm.ed.ac.uk.
7
The Wellcome Trust Sanger Institute, Cambridge, CB10 1SA, UK. tamir.chandra@igmm.ed.ac.uk.
8
The Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, EH4 2XU, UK. tamir.chandra@igmm.ed.ac.uk.

Abstract

DNA modifications, especially methylation, are known to play a crucial part in many regulatory processes in the cell. Recently, 5-hydroxymethylcytosine (5hmC) was discovered, a DNA modification derived as an intermediate of 5-methylcytosine (5mC) oxidation. Efforts to gain insights into function of this DNA modification are underway and several methods were recently described to assess 5hmC levels using sequencing approaches. Here we integrate adaptation based multiplexing and high-efficiency library prep into the oxidative Bisulfite Sequencing (oxBS-seq) workflow reducing the starting amount and cost per sample to identify 5hmC levels genome-wide.

KEYWORDS:

5-Hydroxymethylcytosine; DNA; Methylation; Multiplexing; Oxidative bisulfite sequencing; Pooling

PMID:
29224169
DOI:
10.1007/978-1-4939-7481-8_34
[Indexed for MEDLINE]
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