Single-cell Damagenome Profiling by Linear Copying and Splitting based Whole Genome Amplification (LCS-WGA)

Spontaneous DNA damage frequently occurs on the human genome, and it could alter gene expression by inducing mutagenesis or epigenetic changes. Therefore, it is highly desired to profile DNA damage distribution on the human genome and identify the genes that are prone to DNA damage. Here, we present...

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Bibliographic Details
Published inBio-protocol Vol. 12; no. 6; p. e4357
Main Authors Niu, Yichi, Zhu, Qiangyuan, Zong, Chenghang
Format Journal Article
LanguageEnglish
Published United States Bio-Protocol 20.03.2022
Bio-protocol LLC
Subjects
Biology
Methods
High-damage genes
Vulnerable genes
DNA damage
Whole genome amplification (WGA)
Linear amplification
Damagenome
Single cell whole genome amplification (scWGA)
Genome vulnerability
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Summary:Spontaneous DNA damage frequently occurs on the human genome, and it could alter gene expression by inducing mutagenesis or epigenetic changes. Therefore, it is highly desired to profile DNA damage distribution on the human genome and identify the genes that are prone to DNA damage. Here, we present a novel single-cell whole-genome amplification method which employs linear-copying followed by a split-amplification scheme, to efficiently remove amplification errors and achieve accurate detection of DNA damage in individual cells. In comparison to previous methods that measure DNA damage, our method uses a next-generation sequencing platform to detect misincorporated bases derived from spontaneous DNA damage with single-cell resolution.
Bibliography:Contributed equally to this work
ISSN:2331-8325
2331-8325
DOI:10.21769/BioProtoc.4357