HS maintenance

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Содержание


  • perform maintenance wash after each run
  • performing maintenance wash on one fc while other fc is running is possible ( but!!!: the software pauses the run after the current imaging or chemistry operation is complete, the fc door will not open before the running flowcell paused and resume when you loaded used flowcell for maintenance wash)
  • the maintenance wash includes three wash steps:
• first wash with millipore water
• second wash with 1N NaOH
• third wash with millipore water
  • running eight 250 ml bottles on rack A or B takes 20 minutes (one Fc without PE-rack)
  • running eight 250 ml bottles and then 15 ml tubes on PE-rack takes 45 minutes (one Fc with PE-rack)


[править] Maintenance wash equipments

Don’t use same bottles for first and final wash, to save bottels, name them and use same bottles for xy maintenance washes.

For one maintenance wash per Flowcell with PE needed:

  • 24 Corning bottles (a 250ml), eight bottles per wash and per flowcell rack
  • 30 Millipore bottles (a 15ml), ten bottles per wash and per flowcell rack
  • Millipore water (at least: 180ml)
  • 1N NaOH (at least: 90ml)



[править] Starting the Maintenance wash


[править] First Water Wash

  1. [  ]  from the start screen select wash | maintenance
  2. [  ]  decide if you are washing paired-end positions or not
  3. [  ]  load the instrument with millipore water:
    » a. load on rack A & B eight 250 ml bottles with at least 5 ml millipore water onto the instrument.
    » b. washing the HiSeq with PE positions, load twenty 15 ml tubes with at least 5 ml milipore water to the PE-reagent rack and load the rack onto the instrument.
  4. [  ]  load one or two used flow cell`s on position A or/and B and select next on the screen
  5. [  ]  in case you loaded a used flow cell, perform a fluidics check:
    » a. select solution 2 (millipore water/wash buffer) from the dropdown list.
    »b. enter the following default values:
    • Volume: 250
    • Aspiration Rate: 250
    • Dispense Rate: 2,000
    » c. select pump.
    » d. visually inspect the flow cell for bubbles passing through the lanes and leaks near the manifolds.
  6. [  ]  put the eight lines for each flow cell from the waste container into a 250ml bottle and fix it with eg parafilm and select next
  7. [  ]  when the wash is complete, measure the delivered volume.
    should be:32 ml for one SBS-rack (A or B) and 72ml for SBS rack and PE-rack each Flowcell
  8. [  ]  empty the measure-container to proceed with second wash or return the tubing lanes to the waste bottle



[править] Second wash using NaOH

  1. [  ]  load the instrument with 1 N NaOH:
    » a. load on rack A&B eight 250 ml bottles with 5 ml NaOH onto the instrument
    » b. washing the HiSeq including PE positions, load twenty 15 ml tubes with 5 ml NaOH to the PE-reagent rack and load the rack onto the instrument and start the NaOH wash.
  2. [  ]  when the NaOH wash is complete, measure the delivered volume.
    should be: 32ml for one SBS-rack (A or B) and 72ml for SBS rack and PE-rack each Flowcell
  3. [  ]  empty the measure-container and proceed immediately with final millipore wash



[править] Final Water Wash

  1. [  ]  load the instrument with millipore water:
    » a. load on rack A & B eight 250 ml bottles with at least 5 ml millipore water onto the instrument.
    » b. washing the HiSeq with PE positions, load twenty 15 ml tubes with at least 5 ml milipore water to the PE-reagent rack and load the rack onto the instrument.
  2. [  ]  select next to start the final water wash
  3. [  ]  when the NaOH wash is finished, measure the delivered volume.
    should be: 32ml for one SBS-rack (A or B) and 72ml for SBS rack and PE-rack each Flowcell
  4. [  ]  dont`t forget to return the tubing lanes from measure-container to the HiSeq-waste bottle





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