Original research article

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

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关键词: 果蝇, 总RNA, 制备


  1. 1.5 ml离心管
  2. 果蝇
  3. TRIzolTM Reagent (InvitrogenTM, catalog number: 15596026)
  4. TURBO DNA-freeTM Kit (InvitrogenTM, catalog number: AM1907)
  5. 异丙醇
  6. 乙醇
  7. DEPC-H2O


  1. 离心机 (Eppendorf)
  2. Thermomixer compact (Eppendorf)



  1. 取200 μl TRIzol置于1.5 ml离心管中,放在冰上,将麻醉的10只果蝇放入冰上的离心管中,将果蝇碾碎,再加入800 μl TRIzol试剂,颠倒混匀。
  2. 室温放置10 min,间歇混匀数次。
  3. 4 °C,12,000 × g离心10 min,去掉果蝇残体,取上清至新管。
  4. 加入200 μl氯仿,混匀,室温放置5 min。
  5. 4 °C,12,000 × g离心15 min。
  6. 取上清 (约500 μl) 到新的RNAase free的离心管中,加入等体积的异丙醇混匀,室温放置10 min。
  7. 4 °C,12,000 × g离心30 min。
  8. 弃上清,加入500 μl 70%乙醇,颠倒数次,使RNA漂浮。
  9. 4 °C,7,500 × g离心5 min。
  10. 弃乙醇,如果残留的乙醇有挂壁现象,倒掉液体后再短暂离心,将残液用移液器吸出。室温放置直到白色沉淀呈透明装时加入50 μl DEPC-H2O溶解。
  11. 50 °C,600 rpm金属浴震荡混匀10 min或者时不时轻弹混匀。
  12. 加入1 μl DNase,5 μl DNase Buffer,37 °C温育30 min。
  13. 加5 μl DNase inactive reagent,室温放置5 min,间歇混匀2~3次。
  14. 4 °C,10,000 × g离心1.5 min。
  15. 转移上清至新的EP管中,测浓度;-80 °C保存。

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Copyright: © 2019 The Authors; exclusive licensee Bio-protocol LLC.
引用格式:邓娟, 刘南. (2019). 果蝇总RNA制备. Bio-101: e1010254. DOI: 10.21769/BioProtoc.1010254.
How to cite: Deng, J and Liu, N. (2019). Preparation of Drosophila Total RNA. Bio-101: e1010254. DOI: 10.21769/BioProtoc.1010254.

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