Original research article

A brief version of this protocol appeared in:
Rice Protocol e-book

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关键词: 胚乳, RNA, 抽提


  1. RNase free离心管
  2. RNase free枪头
  3. 水稻种子
  4. 液氮
  5. 无水乙醇
  6. 苯酚 (上海生工分装进口试剂,Tris-HCl饱和,pH 7.9)
  7. 氯仿 (国药集团)
  8. NaAC (Sigma, catalog number: 25022)
  9. 尿素 (Sigma, catalog number: U1250)
  10. 盐酸胍 (Sigma, catalog number: G4505)
  11. EDTA (Sigma, catalog number: E5134)
  12. HEPES (pH 7.0) (Sigma, catalog number: H7006)
  13. 抽提缓冲液 (见溶液配方)
  14. 3 M NaAC (pH 5.2) (见溶液配方)


  1. 冷冻离心机
  2. 振荡器
  3. 超净工作台


  1. 在RNase free的离心管中加入600 μl抽提缓冲液,放在冰上预冷。
  2. 将胚乳放在液氮中研磨成粉末,取适量加入到抽提缓冲液中 (视频1)。
  3. 剧烈振荡 (同抽提普通组织RNA,非振荡器上混匀) 10秒 (视频1)。
  4. 快速加入400 μl苯酚和200 μl氯仿,剧烈振荡 (同抽提普通组织RNA,非振荡器上混匀) 后放在冰上10-30分钟 (视频1)。

    视频1. 水稻胚乳样品研磨及RNA的抽提

  5. 12,000 rpm冷冻离心30分钟。
  6. 转移上清至新的离心管中,加入等体积的无水乙醇,混匀后置于-20度1-2小时。
  7. 12,000 rpm冷冻离心15分钟。
  8. 弃上清,用75%乙醇洗涤后超净台吹干,加入500 μl DEPC水溶解。
  9. 加入250 μl苯酚混匀后再加入250 μl氯仿,剧烈振荡。
  10. 12,000 rpm冷冻离心20分钟。
  11. 转移上清 (400 μl左右) 至新的离心管,加入1/10体积的3 M NaAC(pH 5.2)和2倍体积的无水乙醇,-20度放置1小时以上 (或者-70度30分钟以上)。
  12. 12,000 rpm冷冻离心20分钟,75%乙醇洗涤后超净台吹干,加入适量DEPC水溶解。


  1. 抽提缓冲液

  2. 3 M NaAC (pH 5.2)


操作要求与普通RNA抽提相同。该方法改造自Yang et al. (2006)中马铃薯RNA的抽提方法。


  1. Yang, J. W., Song, B. T., Li, Y. J. and Liu, J. (2006). A simple and efficient method for RNA extraction from patato tuber. Journal of Agricultral Biotechnology 14: 297-298.
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Copyright: © 2018 The Authors; exclusive licensee Bio-protocol LLC.
引用格式:陈香嵩, 程赛凤, 王文韬, 周少立, 赵毓. (2018). 水稻胚乳(种子)RNA的抽提. Bio-101: e1010113. DOI: 10.21769/BioProtoc.1010113.
How to cite: Chen, X. C., Cheng, S. F., Wang, W. T., Zhou, S. L. and Zhao, Y. (2018). Extraction of Total RNA from Rice Endosperm. Bio-101: e1010113. DOI: 10.21769/BioProtoc.1010113.

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