JoVE Journal

Biology

Author Produced

This content is Open Access.

Genome-wide Purification of Extrachromosomal Circular DNA from Eukaryotic Cells

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14:26 min

April 4th, 2016

April 4th, 2016

24,751 Views

0:00

Title

1:07

Part 1. Cell Harvesting

1:49

Part 2. Column Purification

4:03

Part 3. Linear DNA Digestion

6:16

Part 4. Amplification of DNA

7:06

Part 5. Sequencing

7:53

Part 6. Representative Results

11:36

Conclusions & Perspectives

文字起こし

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.

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