DNA Fragmentation Analysis
DNA片段化分析
发布: 2014年08月05日第4卷第15期 DOI: 10.21769/BioProtoc.1203 浏览次数: 17576
评审: Fang XuZhaohui LiuRu Zhang
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参见作者原研究论文
The authors used this protocol in:
Apr 2013
Abstract
DNA fragmentation with length corresponding to multiple integer of approximately 180 base pairs is a distinct feature of apoptosis in animals and programmed cell death in plants. This feature can simply be detected by DNA gel electrophoresis followed by ethidium bromide staining, although in some cases it is difficult to distinguish the DNA laddering. We herein describe a protocol to detect a programmed cell death-associated DNA laddering of plant tissues. After agarose-gel electrophoresis of genomic DNA, Southern hybridization using DIG-labeled genomic DNA probe is performed, that improves detection of DNA laddering.
Keywords: Programmed cell death (程序性细胞死亡)
DNA laddering (DNA梯)
Southern hybridization (Southern杂交)
Materials and Reagents
- Fresh plant tissues
- Nucleon PhytoPure DNA extraction kits (General Electric Company, catalog number: RPN8510 )
- GeneMate LE Agarose (BioExpress, catalog number: E-3120 )
- Biodyne Plus Nylon Membrane (Pall, catalog number: 60406 )
- Wizard DNA Clean-Up System (Promega, catalog number: A7280 )
- DIG-High Prime DNA Labeling and Detection Starter Kit II (Roche Diagnostics, catalog number: 11585614910 )
- X-ray film
- Saline sodium citrate (SSC)
- Depurination solution (see Recipes)
- Denaturation solution (see Recipes)
- Neutralization solution (see Recipes)
- 1x TAE buffer (see Recipes)
Equipment
- Plastic container for gel and membrane treatment
- Centrifuge (Eppendorf, model: 5415R )
- Spectrophotometer (Shimadzu, model: BioSpec-nano )
- Shaker (BIO CRAFT, model: BC-700 )
- Vacuum transfer apparatus (BIO CRAFT, model: BS-31 )
- Hybridization oven and UV crosslinker (UVP, model: HybriLinker HL-2000 )
- Vacuum concentrator (SavantTM, model: DNA SpeedVac DNA110 )
- Hybridization incubator (TAITEC, model: HB-80 )
Procedure
文章信息
版权信息
© 2014 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Mishiba, K., Nagashima, Y., Hayashi, N. and Koizumi, N. (2014). DNA Fragmentation Analysis. Bio-protocol 4(15): e1203. DOI: 10.21769/BioProtoc.1203.
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分类
植物科学 > 植物分子生物学 > DNA
分子生物学 > DNA > DNA 损伤和修复
分子生物学 > DNA > 电泳
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