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A Simple RNA Preparation Procedure from Yeast for Northern Blot Using Hot Phenol

用于Northern印记杂交实验的酵母RNA的提取-简易热苯酚法

Yuehua Wei Yuehua Wei

发布: 2012年06月20日第2卷第12期 DOI: 10.21769/BioProtoc.209 浏览次数: 14776

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Abstract

Compared to several expensive RNA extraction kits, the following protocol provides an economic and simple method for researchers to extract yeast RNA. This method can achieve RNA quality that is sufficient for most northern blot studies in yeast.

Materials and Reagents

  1. W303a cell line
  2. Sodium acetate trihydrate (CH3CO2Na·3H2O) (Sigma-Aldrich, catalog number: 236500 )
  3. Phenol (C6H5OH) (Sigma-Aldrich, catalog number: P1037 )
  4. EDTA (Na2EDTA·2H2O) (Sigma-Aldrich, catalog number: ED2SS )
  5. Acetic acid (CH3COOH) (Sigma-Aldrich, catalog number: 320099 )
  6. Chloroform (CHCl3) (Sigma-Aldrich, catalog number: 472476 )
  7. 8-hydroxyquinoline (C9H7NO) (Sigma-Aldrich, catalog number: 252565 )
  8. NaCl (Thermo Fisher Scientific, catalog number: S641-500 )
  9. Synthetic complete (SC) medium
  10. SDS (Sigma-Aldrich, catalog number: L3771 )
  11. Isoamyl alcohol (Sigma-Aldrich, catalog number: W205710 )
  12. Ethanol (Thermo Fisher Scientific, catalog number: 64-17-5 )
  13. DEPC water
  14. Phenol-AE buffer (see Recipes)
  15. Phenol: CHCl3/AE-Na (see Recipes)
  16. CHCl3: Isoamyl alcohol (24:1) (see Recipes)

Equipment

  1. Standard bench-top centrifuge
  2. Microfuge
  3. Shaker
  4. 1.5 eppendorf tube
  5. Liquid nitrogen

Procedure

English
中文翻译

文章信息

版权信息

© 2012 The Authors; exclusive licensee Bio-protocol LLC.

如何引用

Wei, Y. (2012). A Simple RNA Preparation Procedure from Yeast for Northern Blot Using Hot Phenol. Bio-protocol 2(12): e209. DOI: 10.21769/BioProtoc.209.

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分类

分子生物学 > RNA > RNA 提取

微生物学 > 微生物遗传学 > RNA > RNA 提取

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