期刊
MITOCHONDRION
卷 61, 期 -, 页码 179-187出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.mito.2021.10.005
关键词
Mitochondrial DNA; mtDNA; DNA integrity; Long-range PCR; Nuclease activity
资金
- Swedish Research Council [2019-01874]
- Swedish Cancer Society [19 0022, 190098 Pj 01]
- Swedish Society for Medical Research [S17-0023]
- Ake Wiberg Foundation [(M20-0132)]
This study demonstrates that commonly-used isolation procedures for mitochondrial DNA (mtDNA) result in mtDNA with single-strand breaks. By comparing different DNA isolation methods, a procedure is identified that yields the highest integrity of mtDNA and shows improved performance in downstream assays. These findings emphasize the importance of choosing the right isolation method for obtaining high-quality mtDNA from solid tissues.
The integrity of mitochondrial DNA (mtDNA) isolated from solid tissues is critical for analyses such as long-range PCR, but is typically assessed under conditions that fail to provide information on the individual mtDNA strands. Using denaturing gel electrophoresis, we show that commonly-used isolation procedures generate mtDNA containing several single-strand breaks per strand. Through systematic comparison of DNA isolation methods, we identify a procedure yielding the highest integrity of mtDNA that we demonstrate displays improved performance in downstream assays. Our results highlight the importance of isolation method choice, and serve as a resource to researchers requiring high-quality mtDNA from solid tissues.
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