Prepare

• 5% DECON for washing (~40ml);
• 100ml glass beaker with 5% DECON (Fig.1);
• empty 100ml glass beaker for rinsing ends of tubes (Fig.2);
• set of water-wash-tubes (WWT), without marks:
  • 7x 50ml tubes with 40ml H₂O;
  • 2x 15ml tubes with 10ml H₂O;
  • 9x 1.7ml screw-cup tubes with 1,5ml H₂O;
• set of new water-storage-tubes (WST), with numbers:
  • 7x 50ml tubes with 40ml H₂O: NN 1,2,9,10,11,12,13;
  • 2x 15ml tubes with 10ml H₂O: NN 3,4;
  • 9x 1.7ml screw-cup tubes with 1.5ml H₂O: NN : 5,6,7,8,14,15,16,17,18;

Protocol

Fig. 1.  DECON wash.

1. turn on CS, 30 s, turn on computer, open the program;
2. clean with a wet paper towel:
   • CS inside (except flowcell area),
   • CS outside,
   • the table;
3. empty waste container, rinse it with water;
4. cleaning flowcell area:
   • air [26 · 240µl/min · 160µl],
   • remove bridge manifold,
   • clean with lens cleaning tissue:
          (i) bridge manifold connection sites,
          (ii) quick connect sites,
          (iii) flowcell stage;
   • reconnect bridge manifold;
5. check for leakage:
   • pump water [2 · 240µl/min · 160µl],
   • check that the flow rate in all channels is similar (if not, go to step 4);
   • check for leakage with a folded lens cleaning tissue (if some, go to step 4);
6. DECON wash:
   • unscrew all 1.7ml-caps,
   • prepare glass beaker with 5% DECON (~4cm depth),
   
   
   sequentially for each CS positions (X=1÷18):
   • remove the water-storage-tube (WST),
   • put the tubing (~4cm) into 5% DECON (Fig.1) and wash it [X · 240µl/min · 240µl],
   • remove 5% DECON,
   • air gap [X · 240µl/min · 10µl],
   • rinse the end of the tubing using washing bottle (Fig.2),
   • air gap [X · 240µl/min · 10µl],
   • put the tubing into correspondent WWT (discard old WST tube);

Fig. 2.  Rinsing the end of the tube.

7. first water wash (~15min)
   • recipe window: "DECON_Wash_All_lines_v3.0.xml", "MaintenanceWashDeconWater", "Start", "OK";
   
   wash order:
      240µl: 1,2,3,4,9,10,11,12,13
      120µl: 5,6,7,8,14,15,16,17,18
   
   • during wash change water in WWT's: rinse them and fill with fresh mQ;
8. second water wash (~15min): recipe window as in previous step;


9. substitute WWT tubes with new WST's;


10. close the program; turn off computer, turn off CS;
11. discard WWT and old WST tubes;
12. make a note in the washing calendar.

Notes

DECON 90:
• alkaline solution (with some detergent?), should be stored in a tightly closed flask w/o contact with polycarbonate (borosilicate glass bottle is acceptable);
• ordering:
  
  • England: Decon Laboratories Limited;
  • Germany: Zinsser Analitic: decon90, 3x1L — minimal order, 1L — 12,40€, tel: +49 69 789106/0;


Washing:
• takes ~1hour;
• take care to avoid contact of DECON with caps and tubing of Cluster Station (CS);