用于Northern印记杂交实验的酵母RNA的提取-简易热苯酚法

Yuehua Wei

doi:10.21769/BioProtoc.209

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Peer-reviewed

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 email

发布: 2012年06月20日第2卷第12期 DOI: 10.21769/BioProtoc.209 浏览次数: 14148

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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

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

Equipment

Standard bench-top centrifuge
Microfuge
Shaker
1.5 eppendorf tube
Liquid nitrogen

Procedure

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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.