| Chapter title |
In Situ Analysis of Mitochondrial DNA Synthesis Using Metabolic Labeling Coupled to Fluorescence Microscopy.
|
|---|---|
| Chapter number | 8 |
| Book title |
Mitochondrial DNA
|
| Published in |
Methods in molecular biology, January 2023
|
| DOI | 10.1007/978-1-0716-2922-2_8 |
| Pubmed ID | |
| Book ISBNs |
978-1-07-162921-5, 978-1-07-162922-2
|
| Authors |
Smolka, John A, Lewis, Samantha C, Smolka, John A., Lewis, Samantha C. |
| Abstract |
Metabolic labeling with the nucleoside analog 5-ethynyl-2'-deoxyuridine (EdU) enables the selective labeling of DNA synthesis in live cells. Newly synthesized EdU-containing DNA can be covalently modified after extraction or in fixed cells using copper-catalyzed azide-alkyne cycloaddition "click chemistry" reactions, enabling bioconjugation to various substrates including fluorophores for imaging studies. While often used to study nuclear DNA replication, EdU labeling can also be leveraged to detect the synthesis of organellar DNA in the cytoplasm of Eukaryotic cells. In this chapter, we outline methods for the application of EdU labeling to the study of mitochondrial genome synthesis in fixed cultured human cells, using fluorescent labeling and superresolution light microscopy. |
X Demographics
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| Members of the public | 2 | 100% |
Mendeley readers
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| Readers by discipline | Count | As % |
|---|---|---|
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| Unknown | 2 | 50% |