Materials and Reagents

1. Glass tubes [PYREX^TM screw cap culture tubes with PTFE lined phenolic caps (Thermo Fisher Scientific, Fisher Scientific^TM, catalog number: 14-932H )]
   
2. Pipette 100-1,000 µl (Eppendorf Research 2100)
   
3. Axygen^TM 1,000 µl universal pipette tips (Thermo Fisher Scientific, Fisher Scientific^TM, catalog number: 14-222-690 )
   
4. Total plant lipids
   Note: Extracted by the method described in the companion lipid extraction and profiling protocol by the same authors (Liu and Wang, 2016) and the original publication (Welti et al., 2002)
   
5. Chloroform (Thermo Fisher Scientific, catalog number: C607 )
   
6. PLA₂ from honey bee venom (Sigma-Aldrich, catalog number: P9279 )
   Note: Dissolved in reaction buffer to 1 unit/µl before use.
   
7. Sodium chloride (NaCl) (Sigma-Aldrich, catalog number: S7653 )
   
8. Nitrogen gas
   
9. HEPES (Sigma-Aldrich, catalog number: H3375 )
   
10. Calcium chloride (CaCl₂) (Sigma-Aldrich, catalog number: C8106 )
    
11. Magnesium chloride hexahydrate (MgCl₂·6H₂O) (Sigma-Aldrich, catalog number: M2670 )
    
12. Methanol (Thermo Fisher Scientific, Fisher Scientific^TM, catalog number: A456 )
    
13. Reaction buffer (see Recipes)
    
14. NaCl solution (see Recipes)
    
15. Extraction solution (see Recipes)

Equipment

1. Nitrogen evaporator N-EVAP (Meyer Organomation, model: 111 )
   
2. Ultrasonic processor with probe, work at 30% power (GEX 130PB) or similar equipment [e.g., ultrasonic processor, (Cole-parmer, model: EW-04714-51 )]
   
3. Incubator (Thermo Fisher Scientific, model: Isotemp 537D ) or similar equipment [e.g., microbiological incubators (Thermo Fisher Scientific, Isotemp^TM, catalog number: 15-103-0513 )]

Procedure