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Determination of Mitochondrial DNA Upon Drug Treatment

药物处理后的线粒体DNA测定

MP Michel Perron
Joy Y. Feng Joy Y. Feng

发布: 2015年01月20日第5卷第2期 DOI: 10.21769/BioProtoc.1377 浏览次数: 9041

评审: Vanesa Olivares-IllanaPia GiovannelliAnonymous reviewer(s)

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Abstract

Drug-induced mitochondrial injury can be caused by many different mechanisms including inhibition of mitochondrial DNA replication, transcription, translation, and altered protein function. Determination of the level of mitochondrial DNA relative to the nuclear DNA levels provides important information on potential mitochondrial toxicity.

Keywords: Mitochondrial toxicity (线粒体毒性) search Nucleoside analogs (核甘酸类似物) search Inhibitor of DNA synthesis (DNA合成抑制剂) search

Materials and Reagents

  1. HepG2 cells (ATCC, catalog number: HB 8065 )
  2. DMSO (cell culture grade) (Sigma-Aldrich, catalog number: D2650 )
  3. Phosphor-buffered saline (PBS) (Life Sciences, catalog number: 10010049 )
  4. QIAamp DNA mini kit (QIAGEN, catalog number: 51304 )
  5. TaqMan universal mastermix (Life Technologies, Applied Biosystems®, catalog number: 4352042 )
  6. β-actin Assay-on-Demand kit (Life Technologies, Applied Biosystems®, catalog number: 4331182 )
  7. Eagle’s minimum essential medium (Life Technologies, Gibco®, catalog number: 41090 )
  8. GlutaMAXTM
  9. Fetal bovine serum (FBS) (HyClone, catalog number: SH30071.03 )
  10. 100 units/ml penicillin, 100 units/ml streptomycin (Life Technologies, Gibco®, catalog number: 15140 )
  11. Sodium pyruvate (Life Technologies, Gibco®, catalog number: 11360 )
  12. Cells were cultured in Eagle’s minimum essential medium (see Recipes)
    Note: Cells were cultured in Eagle’s minimum essential medium.

Equipment

  1. 12-well plates (Corning, catalog number: 3513 )
  2. ABI Prism 7900HT Fast Real-Time PCR system (Life Technologies, Applied Biosystems®)

Procedure

English
中文翻译

文章信息

版权信息

© 2015 The Authors; exclusive licensee Bio-protocol LLC.

如何引用

Perron, M. and Feng, J. Y. (2015). Determination of Mitochondrial DNA Upon Drug Treatment. Bio-protocol 5(2): e1377. DOI: 10.21769/BioProtoc.1377.

ris ris Download Citation in RIS Format

分类

分子生物学 > DNA > DNA 损伤和修复

分子生物学 > DNA > DNA 定量

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