Materials and Reagents

1. Guanidinium thiocyanate (Thermo Fisher Scientific, catalog number:  BP221 )
2. NaCitrate
3. N-Lauroylsarcosine (Sigma-Aldrich, catalog number: L9150 )
4. β-mercaptoethanol (β-ME)
5. Glacial HAc
6. NaOH
7. Phenol (J.T.Baker^®, catalog number:  2859 )
8. Ethylenediaminetetraacetic acid (EDTA)
9. Sodium dodecyl sulfate (SDS) (Sigma-Aldrich, catalog number: L5750 )
10. Tris (hydroxymethyl) aminomethane
11. Chloroform
12. Isopropanol
13. Diethylpyrocarbonate (DEPC)
14. Ethanol
    
15. Extraction buffer (EB) (see Recipes)
    
16. 2 M NaAcetate (pH 4.0) (see Recipes)
17. 2 M NaAcetate (pH 5.0) (DEPC treated) (see Recipes)
    
18. H₂O-saturated phenol (no buffer required) (see Recipes)
    
19. Citrate-EDTA-SDS solution (CES) (see Recipes)
    
20. Tris-EDTA-SDS solution (TES) (see Recipes)
    
21. 100 ml 0.75 M NaCitrate (see Recipes)
    
22. 60 ml 10% Sarkosyl (see Recipes)
    

Equipment

1. Vortex Mixer 
2. Beckman high speed centrifuge with JS13.1 rotor or the like
3. 15-ml Corning centrifuge tube
   

Procedure