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Integrated single-cell sequencing of 5-hydroxymethylcytosine and genomic DNA using scH&G-seq.

ABSTRACT: The asymmetric distribution of 5-hydroxymethylcytosine (5hmC) between two DNA strands of a chromosome enables endogenous reconstruction of cellular lineages at an individual-cell-division resolution. Further, when integrated with data on genomic variants to infer clonal lineages, this combinatorial information accurately reconstructs larger lineage trees. Here, we provide a detailed protocol for single-cell 5-hydroxymethylcytosine and genomic DNA sequencing (scH&G-seq) to simultaneously quantify 5hmC and genomic DNA from the same cell to reconstruct lineage trees at a single-cell-division resolution. For complete details on the use and execution of this protocol, please refer to Wangsanuwat et al., 2021.

SUBMITTER: Chialastri A

PROVIDER: S-EPMC8672100 | biostudies-literature | 2021 Dec

REPOSITORIES: biostudies-literature

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Integrated single-cell sequencing of 5-hydroxymethylcytosine and genomic DNA using scH&G-seq.

Chialastri Alex A Wangsanuwat Chatarin C Dey Siddharth S SS

STAR protocols 20211211 4

The asymmetric distribution of 5-hydroxymethylcytosine (5hmC) between two DNA strands of a chromosome enables endogenous reconstruction of cellular lineages at an individual-cell-division resolution. Further, when integrated with data on genomic variants to infer clonal lineages, this combinatorial information accurately reconstructs larger lineage trees. Here, we provide a detailed protocol for single-cell 5-hydroxymethylcytosine and genomic DNA sequencing (scH&G-seq) to simultaneously quantify ...[more]

PMID: 34950891

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