Circle-Seq: Isolation and Sequencing of Chromosome-Derived Circular DNA Elements in Cells
Publikation: Bidrag til bog/antologi/rapport › Bidrag til bog/antologi › Forskning › fagfællebedømt
Chromosome-derived extrachromosomal circular DNA elements (eccDNAs) are detected in all eukaryotes examined so far. Here I describe the Circle-Seq protocol, applicable for physical enrichment of eccDNAs of a broad size range, combined with sequence confirmation of circular structures.Briefly, by concise alkaline treatment and gentle gravity flow-through an ion-exchange column, eccDNAs are enriched in the eluate fraction. EccDNAs are enzymatically isolated by extensive Plasmid-Safe DNase digestion of linear chromosomes and further enriched by φ29 rolling circle amplification. By means of high throughput sequencing of amplified eccDNA and custom eccDNA mapping software, around ten-thousand unique eccDNA types could be detected at nucleotide resolution in a million human muscle nuclei by this method.
Originalsprog | Engelsk |
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Titel | DNA Electrophoresis : Methods and Protocols |
Redaktører | Katsuhiro Hanada |
Antal sider | 17 |
Forlag | Humana Press |
Publikationsdato | 2020 |
Sider | 165-181 |
Kapitel | 15 |
ISBN (Trykt) | 978-1-0716-0322-2 |
ISBN (Elektronisk) | 978-1-0716-0323-9 |
DOI | |
Status | Udgivet - 2020 |
Navn | Methods in Molecular Biology |
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Vol/bind | 2119 |
ISSN | 1064-3745 |
ID: 235588304