HiSeq 2000 general information

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General info

  1. FC compartment
    • the HiSeq 2000 (HS) can run two FC`s at same time with output ~200GB mappable data per run (state June 2011)
    • HS can run two FC`s with different read lengths simultaneously, and start and stop experiments independently.
    • both Fc`s are positioned on the flow cell holder and are hold by vacuum with LED (shows green or yellow when vacuum is on or off),no oil and no prism as in the GAII
    • stage can move in and out of the optics module. Flow cell A is on the left, and flow cell B is on the right side. Each flow cell is with the inlet and outlet holes facing down.
    • other orientation of the lines as on the cBot

  2. FC
    • HS-FC ist lager (5x imaging area) than GAII-FC, compatible only with cBot

  3. fluidics
    • two fluidics pumps, one for each flow cell. Pumps direct reagents through the fluidics lines to the flow cell, and then deliver waste to the waste bottle.

  4. reagents
    • two racks for sequencing reagents for 250ml bottles for FC's A&B and one PE-Rack for paired-end and multiplexing reagents with two rows, one for each FC
    • temperature-controlled (high-capacity chiller) but not for cooling down the reagents (reagents must be cooled before loading to the instrument)
    • enough capacity for sequencing reagents up to 209cycles per Fc
    • reagents are bar-coded for tracking

  5. imaging
    • images A, C, G, and T at the same time using a four camera epi-fluorescence system. EPI-fluoescence: the excitation laser beam passes through the objective and the fluorescence is collected through the same objective.
    • dual surface imaging

  6. software
    • interface provides dual controls, one for each flow cell, on a touch screen enabled monitor. In addition, an integrated keyboard can be used for recording run components and operating the instrument.
    • real time data analysis on the instrument computer, which allows to monitor important quality metrics during the
    sequencing run


  • Instrument status: Blue -- instrument is running, orange -- instrument needs attention, green -- instrument is ready to begin the next run.


to be done: What is happening on HiSeq?

Working place near the HiSeq

  • used FC
  • cleaning tissue for the FC (Whatman)
  • new gaskets
  • washing bottle with mQ
  • dust-free paper towels
  • plastic forceps
  • 15 ml tubes, one line per tube


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