Materials and Reagents

1. 15 ml Falcon tubes (VWR International, catalog number: 734-0452 )
   
2. 2 ml Eppendorf safe-lock tubes (VWR International, catalog number: 211-2165 )
   
3. 2 ml screw-cap tubes (VWR International, catalog number: 211-0093 )
   
4. Chromatography vials with inserts (VWR International, catalog number: 548-0120 )
   
5. Snap cap for chromatography vials (VWR International, catalog number: 548-1151 )
   
6. Manual pipettes tips
   
7. Arabidopsis thaliana seeds
   
8. 2-Deoxy-D-glucose (2-deoxy-Glc) (Sigma-Aldrich, catalog number: D6134-1 G )
   
9. D-(-)-Ribose (Rib) (Sigma-Aldrich, catalog number: R7500-5 G )
   
10. L-(+)-Arabinose (Ara) (Sigma-Aldrich, catalog number: A3256-25 G )
    
11. L-(-)-Fucose (Fuc) (Sigma-Aldrich, catalog number: F2252-5 G )
    
12. D-(+)-Galactose (Gal) (Sigma-Aldrich, catalog number: G0750-25 G )
    
13. D-(+)-Galacturonic acid monohydrate (GalA) (Sigma-Aldrich, catalog number: 48280-5G-F )
    
14. D-(+)-Glucose (Glc) (Sigma-Aldrich, catalog number: G8270-100 G )
    
15. D-Glucuronic acid (GlcA) (Sigma-Aldrich, catalog number: G5269-10 G )
    
16. D-(+)-Mannose (Man) (Sigma-Aldrich, catalog number: M8574-25 G )
    
17. L-Rhamnose monohydrate (Rha) (Sigma-Aldrich, catalog number: R3875-5 G )
    
18. D-(+)-Xylose (Xyl) (Sigma-Aldrich, catalog number: X3877-25 G )
    
19. Ultrapure water (18.2 MΩ cm at 25 °C)
    
20. Sodium hydroxide (NaOH) (VWR International, catalog number: BAKR3727.2500 )
    
21. Trifluoroacetic acid (TFA) (Carl Roth GmbH + Co., catalog number: 6957.1 )
    
22. Sugar standard stocks (see Recipes)
    
23. 9-Sugar mix (see Recipes)
    
24. 2 M TFA (see Recipes)
    
25. 10 mM NaOH (see Recipes)
    
26. 733 mM NaOH (see Recipes)
    

Equipment

1. Autoclave
   
2. Water purification system (Milli-Q or similar style)
   
3. Manual pipettes (Eppendorf AG, Research plus and Repeater Plus style)
   
4. Analytical balance (Mettler-Toledo, model: XSE205DU )
   
5. Ball mill (Retsch GmbH, catalog number: MM400 )
   
6. Two 24 TissueLyser adapters for ball mill (QIAGEN, catalog number: 69982 )
   
7. Lab coat
   
8. Safety glasses
   
9. Chemical resistant gloves (Honeywell, Dermatril, catalog number: 740 , or similar style)
   
10. Fume hood
    
11. Sample concentrator (Bibby Scientific Limited, Techne, catalog number: FSC400D ), equipped with
    1. 127 mm needles (Bibby Scientific Limited, Techne, catalog number: F7210 )
       
    2. A Dri-block heater (Bibby Scientific Limited, Techne, catalog number: DB200/3 )
       
    3. Three aluminium blocks (Bibby Scientific Limited, Techne, catalog number: F3505 )
       
    4. Connected to a central air supply, in a fume hood
       
12. Ice machine
    
13. Ion chromatography (IC): Dionex DX-600 system equipped with
    1. AS50 autosampler
       
    2. GP50 gradient pump
       
    3. ED50 electrochemical detector
       
    4. CarboPac PA20 guard column (Thermo Fisher Scientific, Dionex Softron, catalog number: 060144 )
       
    5. CarboPac PA20 analytical column (Thermo Fisher Scientific, Dionex Softron, catalog number: 60142 )
       
14. Vortex mixer (Scientific Industries Inc., model: Vortex-Genie 2 , or similar style)
    
15. Benchtop centrifuge (compatible with 2 ml tubes)
    
16. Racks with lids for 2 ml tubes (VWR International, catalog number: 211-0215 )
    
17. Serological pipettes (Thermo Fisher Scientific, Nunc-type or similar style)
    
18. 2 L volumetric flask
    
19. Helium gas tank
    

Software

1. Chromeleon 6.8 chromatography data system software (Thermo Fisher Scientific, Dionex)
   
2. Microsoft Excel with the Real Statistics Resource Pack (http://www.real-statistics.com/)
   

Procedure