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一种简易可行的激光捕获显微切割样本RNA质控方法 被引量：1

A simple method for assessment of RNA integrity in laser capture microdissection samples
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摘要 目的建立一种简易可行的激光捕获显微切割样本RNA质控方法。方法分别提取捕获样本和染色前、染色后1和2h、持续捕获1和2h切片及残余组织的RNA,琼脂糖电泳检测;激光捕获显微切割(LCM)样本RNA的产率可通过理论值初步估计。结果当染色前后、捕获后的切片或切片残余组织RNA质量完好时,捕获样本的RNA质量完好,实时定量PCR亦获得了良好的扩增结果。结论使用琼脂糖凝胶电泳分别检测染色前后、捕获后的切片或切片残余组织RNA的质量,可以从不同的环节和过程对捕获样本RNA进行间接检测,该方法简单方便有效,并可进一步用于LCM样本蛋白质及DNA等大分子的质控。 Objective To develop a simple method for assessment of RNA integrity in laser capture microdissection （LCM） samples. Methods The total RNA were isolated from the LCM samples and the sections before and after microdissection and examined by agarose gel electrophoresis. Real-time PCR was employed to assess the RNA from LCM samples, and the quantity of RNA was theoretically estimated according to the average total RNA product in mammalian cells （10 ng/1000 cells）. Results When the total RNA from the sections before and after microdissection was intact, the RNA from LCM samples also had good quality, and the 28S and 18S rRNAs were visualized by ethidium bromide staining. Real-time PCR also showed good RNA quality in the LCM samples. Conclusion A simple method for quantitative and qualitative assessment of the RNA from LCM samples is established, which can also be applied to assessment of DNA or proteins in LCM samples.
作者 田英芳 魏朝明 陈新林 邱芬 肖新莉 康前雁 朱波峰 田玉梅 张军峰 刘勇
机构地区 西安交通大学医学院环境与疾病相关基因教育部重点实验室/神经生物学研究所 西安交通大学医学院法医学国家重点学科 西安交通大学医学院生理学国家重点学科
出处 《南方医科大学学报》 CAS CSCD 北大核心 2008年第10期1782-1785,共4页 Journal of Southern Medical University
基金 国家自然科学基金(30500575) 陕西省科技攻关项目(2006K13-G5)
关键词 激光捕获显微切割 RNA质控 琼脂糖凝胶电泳 laser capture microdissection RNA integrity agarose gel electrophoresis
分类号 Q522 [生物学—生物化学]
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参考文献10

  • 1Simone NL, Bonner RF, Gillespie JW, et al. Laser-capture microdissection: opening the microscopic frontier to molecular analysis [J ]. Trends Genet, 1998, 14(7): 272-6.
  • 2Burgess JK, Hazelton RH. New developments in the analysis of gene expression [J]. Redox Rep, 2000, 5(2): 63-73.
  • 3Vincent VA, DeVoss JJ, Ryan HS, et al. Analysis of neuronal gene expression with laser capture microdissection [J]. J Neurosci Res, 2002, 69(5): 578-86.
  • 4Shibutani M, Uneyama C, Miyazaki, et al. Methacam: a fixation tool for multipurpose genetic analysis from paraffin-embedded tissues [J]. Methods Enzymol, 2002, 356:114-25.
  • 5Luzzi V, Mahadevappa M, Raja R, et al. Accurate and reproducible gene expression profiles from laser capture microdissection, transcript amplification, and high density oligonucleotide microarray analysis [J]. J Mol Diagn, 2003, 5(1): 9-14.
  • 6Betsuyaku T, Senior RM. Laser capture microdissection and mRNA characterization of mouse airway epithelium: methodological considerations[J]. Micron, 2004, 35(4): 229-34.
  • 7田英芳,刘勇,张蓬勃.激光捕获显微切割技术及其在脑研究中的应用[J].解剖科学进展,2005,11(2):170-173. 被引量：4
  • 8田英芳,魏朝明,张蓬勃,邱芬,陈新林,刘勇.激光显微切割联合功能分类表达谱芯片技术在脑缺血研究中的应用[J].西安交通大学学报（医学版）,2005,26(4):395-398. 被引量：3
  • 9Shimamura M, Garcia JM, Prough DS, et al. Laser capture microdissection and analysis of amplified antisense RNA from distinct cell populations of the young and aged rat brain: effect of traumatic brain injury on hippocampal gene expression [J].Brain Res Mol Brain Res, 2004, 122(1): 47-61.
  • 10Keays KM, Owens GP, Ritchie AM, et al. Laser capture microdissection and single-cell RT-PCR without RNA purification [J].J Immunol Methods, 2005, 302(1-2): 90-8.

二级参考文献33

  • 1Vincent VA, De Voss JJ, Ryan HS, et al. Analysis of neuronal gene expression with laser capture microdissection [ J ]. J Neurosci Res,2002 ,69(5) :578 -586.
  • 2Robles Y, Vivas - Mejia PE, Ortiz-Zuazaga HG, et al. Hippocampal gene expression profiling in spatial discrimination learning [J]. Neurobiol Learn Mem, 2003, 80(1): 80-95.
  • 3Bonaventure P, Guo H, Tian B, et al. Nuclei and subnuclei gene expression profiling in mammalian brain [ J ]. Brain Res, 2002,943(1 ): 38 -47.
  • 4Lu L, Neff F, Dun Z, et al. Gene expression profiles derived from single cells in human postmortem brain[J]. Brain Res Brain Res Protoc, 2004,13(1): 18 -25.
  • 5Kamme F, Zhu J, Luo L, et al. Single-cell laser-capture microdissection and RNA amplification [ J ]. Methods Mol Med,2004, 99:215-224.
  • 6Mouledous L, Hunt S, Harcourt R, et al. Proteomic analysis of immunostained, laser - capture microdissected brain samples[ J ].Electrophoresis, 2003, 24 ( 1 - 2): 296 - 302.
  • 7Mouledous L, Hunt S, Harcourt R, et al. Navigated laser capture microdissection as an alternative to direct histological staining for proteomic analysis of brain samples [ J ]. Proteomics. 2003, 3(5): 610 -615.
  • 8Liao L, Cheng D, Wang J, Proteomic characterization of postmortem amyloid plaques isolated by laser capture microdissection[ J].J Biol Chem, 2004, 279(35) :37061 -37068.
  • 9Mojsilovic-Petrovic J, Nesic M, Pen A, et al. Development of rapid staining protocols for laser - capture microdissection of brain vessels from human and rat coupled to gene expression analyses [J]. J Neurosci Methods, 2004,133(1 -2) :39 -48.
  • 10Parlato R, Rosica A, Cuccurullo V, et al. A preservation method that allows recovery of intact RNA from tissues dissected by Laser capture microdissection[J]. Anal Biochem, 2002, 300(2): 139-145.

共引文献5

同被引文献17

  • 1Vincent VA, DeVoss JJ, Ryan HS, et al. Analysis of neuronal gene expression with laser capture microdissection. J Neurosci Res, 2002, 69(5) : 578-586.
  • 2Wang WZ, Oeschger FM, Lee S, et al. High quality RNA from multiple brain regions simultaneously acquired by laser capture microdissection. BMC Mol Biol, 2009, 10: 69.
  • 3Emmert-Buck MR, Bonner RF, Srsith PD, et al. Laser capture microdissection. Science(80-), 1996,274(5289) : 998-1001.
  • 4邱宗荫,尹一兵.临床蛋白质组学.1版.北京:科学出版社,2008.
  • 5Mojsilovic-Petrovic J, Nesic M, Pen A, et al. Development of rapid staining protocols for laser-capture microdissection of brain vessels from human and rat coupled to gene expression analyses. J Neurosci Methods, 2004, 133(1-2): 39-48.
  • 6Hellmich HL, Capra B, Eidson K, et al. Dose-dependent neuronal injury after traumatic brain injury. Brain Res, 2005, 1044(2): 144- 154.
  • 7Liu Y, Wu J, Liu S, et al. Immuno-laser capture microdissection of frozen prolactioma sections to prepare proteomic samples. Colloids Surf B Biointerfaces, 2009, 71(2): 187-193.
  • 8Markaryan A, Nelson EG, Hinojosa R. Detection of mitochondrial DNA deletions in the cochlea and its structural elements from archival human temporal bone tissue. Mutat Res, 2008, 640 (1-2) : 38-45.
  • 9Harris LW, Wayland M, Lan M, et al. The cerebral microvasculature in schizophrenia: a laser capture microdissection study. PLoS One, 2008, 3(12) : e3964.
  • 10Kamme F, Salunga R, Yu J, et al. Single-cell microarray analysis in hippocampus CA1 : demonstration and validation of cellular heterogeneity. J Neurosci, 2003, 23(9): 3607-3615.

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南方医科大学学报
2008年 第10期

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