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Chapters
- 00:00Title
- 01:07Part 1. Cell Harvesting
- 01:49Part 2. Column Purification
- 04:03Part 3. Linear DNA Digestion
- 06:16Part 4. Amplification of DNA
- 07:06Part 5. Sequencing
- 07:53Part 6. Representative Results
- 11:36Conclusions & Perspectives
This paper presents a sensitive method called Circle-Seq for purifying extrachromosomal circular DNA (eccDNA). The method encompasses column purification, removal of remaining linear chromosomal DNA, rolling-circle amplification and high-throughput sequencing. Circle-Seq is applicable to genome-scale screening of eukaryotic eccDNA and studying genome instability and copy-number variation.
Tags
Keywords:
Extrachromosomal Circular DNAGenome-wide PurificationEukaryotic CellsYeast CellsColumn SeparationExonuclease TreatmentPhi-29 Polymerase AmplificationSaccharomyces CerevisiaeCell HarvestingCell Wall DisruptionColumn PurificationSpiked-in PlasmidsInternal ReferenceSemi-quantitative Estimation
