Materials and Reagents

1. Bodicon m7G capping system (Bodicon, catalog number: CS0130 )
   
2. S-adenosyl methionine (SAM) (involved in Bodicon m7G capping system) (Bodicon, catalog number: CS0130)
   
3. Bodicon Capping Enzyme (10 U/µl) (involved in Bodicon m7G capping system) (Bodicon, catalog number: CS0130)
   Note: Because the sale of this kit was low, the previous companies which provided this capping kit were out of service. This capping kit was provided by a new company in China as custom-made products (contact e-mail: service@bodicon.cn, phone:+86-13628662011). In fact the similar capping kit from any other companies (such as EPICENTRE biotechnologies, ScriptCap m7G capping system, catalog number: SCCE0610 ) is suitable for this experiment, and people can also contact with us to get the related protein or kit.
   
4. Inorganic pyrophospatase (YIPP) (New England Biolabs, catalog number: M2403S )
   
5. S-adenosyl [methyl-³H] methionine ([³H]-SAM) (PerkinElmer, catalog number: NET155H001MC )
   
6. DEAE Sephadex (GE Healthcare, catalog number: 17-0170-01 )
   
7. GTP (Thomas Scientific, catalog number: R0461 )
   
8. RNase inhibitor (Thomas Scientific, catalog number: EO0381 )
   
9. RNase free water
   
10. Phenol-chloroform (pH 4.8-5.2 for RNA only)
    
11. Ethanol (RNase free)
    
12. RNase free water
    
13. Sodium Dodecyl Sulfonate (SDS)
    
14. Ethylene Diamine Tetraacetic Acid (EDTA)
    
15. NH₄HCO₃
    
16. NaCl
    
17. 10x MTase Buffer (see Recipes)
    
18. Cap-0 cap structure (m7GpppN-RNA) (see Recipes)
    
19. Non-methylated Cap-0 cap structure (GpppN-RNA) (see Recipes)
    
20. MTase assay reaction mix (see Recipes)
    

Equipment

1. Bechtop
   
2. Water bath
   
3. Centrifuge
   
4. Liquid scintillation counter
   

Procedure