DECON wash

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NB! post-PCR operation
should be performed once in 2 weeks


Содержание


[править] Prepare

    • 5% DECON for washing (~40ml);
    • 100ml glass beaker with 5% DECON (Fig.1);
    Fig. 1. DECON wash.
    • 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 15 ml tubes with 10ml H2O: NN 3, 4;
      • 9x 1.7ml screw-cup tubes with 1.5ml H2O: 5, 6, 7, 8, 14, 15, 16, 17, 18;

[править] Protocol

  1. turn on CS, 30 s, turn on computer, open the program;
  2. clean with a wet paper towel:
    • CS inside (exept 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 laekage with folded lens cleaning tissue (if some, go to step 4);
    Fig. 2. Rinsing the end of the tube.
  6. DECON wash:
    • unscrew all 1.7ml-caps,
    • prepare glass beaker with 5% DECON (~4cm depth),
    perform sequentially for each CS position (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);
  7. first water wash (~15min)
    • recipe window: "DECON_Wash_ All_lines_v7.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
    • after wash change water in WWT's: rinse them and fill with fresh mQ;
  8. second water wash (~15min): recipe window as in previouse step;
  9. substitute WWT tubes with new WST's; discard WWTs
  10. close the program; turn off computer, turn off CS;
  11. make a note in the washing calendar.

[править] Notes

DECON 90:
  • alkaline solution (with some detergent?), shold be stored in 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: +4969789106/0 begin_of_the_skype_highlighting              +4969789106/0      end_of_the_skype_highlighting;
Washing:
  • takes ~ 2 hours. It is possible to be faster if you change water in WWTs during the first water wash and substitute WWTs to new WSTs during the second water wash.
  • take care to avoid contact of Decon with caps and tubing of Cluster Station (CS).
Источник — «http://www.molbiol.ru/wiki/DECON_wash»


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