Hot phenol SDS method

This is a popular method for preparation of RNA from bacterial, archaeal and small eukaryotes (e.g. yeast). The method exploits the property of DNA to go into the phenol phase together with the proteins at pH 5.0. 

Phenol (saturated with acetate buffer) Equipment 

Discard the supernatant and wash the pellet in 1 ml acetate buffer. Transfer to a microfuge tube and centrifuge for 3 min at 4°C. 

Discard the supernatant and resuspend the pellet in 500 pi ice-cold acetate buffer. 

Add 550 pi phenol (prewarmed at 65°C), vortex for 1 min,b and place in a heating block for 5 min at 65°C (vortex briefly every minute).c 

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