Materials and Reagents

1. Pseudomonas syringae (P. syringae) Lz4W strain (Shivaji et al., 1989)
   
2. Escherichia coli (E. coli) DH5alpha strain
   
3. Random primer labelling kit (JONAKI Laboratory, catalog number: LCK 2 )
   Note: Many companies like Roche Diagnostics, Thermo Fisher Scientific, Agilent, Clontech etc. provide similar kit.
   
4. Wizard Genomic DNA Purification Kit (Promega Corporation, catalog number: A1120 )
   
5. (Alpha ³²P)-dATP (JONAKI Laboratory, catalog number: LCP 103 )
   
6. Lysozyme (Roche Diagnostics, catalog number: 10153516103 )
   
7. 10% trichloroacetic acid (TCA) (Sigma-Aldrich, catalog number: T6399 )
   
8. KCl (Sigma-Aldrich, catalog number: P9541 )
   
9. 50 mM Tris^.HCl (pH 7.5)
   
10. 10% sucrose (Sigma-Aldrich, catalog number: 84097 )
    
11. Peptone (HiMedia Laboratories, catalog number: RM 001-500G )
    
12. Yeast extracts (HiMedia Laboratories, catalog number: RM 027-500G )
    
13. Stop buffer (see Recipes)
    
14. Lysozyme solution (see Recipes)
    
15. ABM broth (Pavankumar et al., 2010) (see Recipes)
    

Equipment

1. BD PrecisionGlide needles 22G (BD, catalog number: 305156 )
   
2. Liquid scintillation analyzer (PerkinElmer, model: Tri-Carb 2900 TR )
   
3. Centrifuge (Eppendorf, model: 5810R, catalog number: 5811000.010 )
   
4. NanoDrop 1000 spectrophotometer (Thermo Fisher Scientific, catalog number 8482 )
   
5. Micro centrifuge tubes (1.5 ml) (tarsons, catalog number: 500010 )
   
6. F12- ED refrigerated/heating circulator (JULABO GmbH, catalog number: 9116612 )
   
7. Plastic scintillation vials (PerkinElmer, catalog number: 6000477 )
   
8. Separate laboratory design and workspace for radioactive work with proper guidelines
   

Procedure