Materials and Reagents

1. Mycobacterium culture (50 to 100 ml)
   
2. Tris base (Sigma-Aldrich, catalog number: T1503 )
   
3. EDTA (HiMedia Laboratories, catalog number: MB011 )
   Note: Prepare 0.5 M solution (pH 8) in double distilled water.
   
4. Liquid nitrogen
   
5. Proteinase K (HiMedia Laboratories, catalog number: RM2957 )
   Note: Prepare the stock solution of 10 mg/ml in TE buffer (pH 8).
   
6. SDS (Bio Basic Canada, catalog number: SB0485 )
   Note: Prepare 10% solution by dissolving in double distilled water.
   
7. Lysozyme (Sigma-Aldrich, catalog number: L6876 )
   Note: Prepare the stock solution of 100 mg/ml in double distilled water.
   
8. Phenol (Sigma-Aldrich, catalog number: P4557 )
   Note: Equilibrate with Tris-Hcl (pH 8).
   
9. Chloroform (HiMedia Laboratories, catalog number: AS041 )
   
10. Isoamyl alcohol (HiMedia Laboratories, catalog number: MB091 )
    
11. 2-Propanol (HiMedia Laboratories, catalog number: MB063 )
    
12. Ethyl alcohol, Pure (Sigma-Aldrich, catalog number: 459844 )
    
13. Nuclease-free water (HiMedia Laboratories, catalog number: TCL016 )
    
14. TE buffer (see Recipes)
    
15. PCI mixture (see Recipes)
    

Equipment

1. Water bath
   
2. Centrifuge
   
3. Pestle and mortar
   

Procedure