Qiagen PyroMark Q96 ID, User Manual

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General Description
3-4 PyroMark Q96 ID User Manual 01/2016
6. Once the temperatures and pressure reach the preset
levels (this may take several minutes), the
predispensation and dispensation of the reagents begins.
7. Enzyme and substrate mixtures are predispensed into the
rectangular sink-well at the right-hand edge of the
PyroMark Q96 Plate Low to ensure that the dispensation
capillaries are flushed and filled with solution.
8. The reagent cartridge is positioned over each well in turn
and the enzyme mixture is dispensed. The process is then
repeated for the substrate mixture.
Note : Dispensation of all reagents into the PyroMark
Q96 Plate Low is in a zigzag fashion. The mixing system
(vibrating process chamber) is operational throughout the
sequencing run.
9. The dispensation pressure is increased.
10. The nucleotides are predispensed into the rectangular
sink-well at the right-hand edge of the PyroMark Q96
Plate Low.
11. The reagent cartridge is positioned over each well in turn
and the programmed nucleotide is dispensed.
12. The CCD camera collects data simultaneously from all
wells throughout the process. The CCD camera is cooled
to approximately 0°C to optimize performance and
reduce background noise.
13. In wells where there is a positive reaction with the added
nucleotide, light is emitted, giving rise to a peak in the
Pyrogram ® .
14. A period of 65 seconds (cycle time) is allowed between
the addition of each nucleotide to ensure that all
enzymatic reactions are completed.
15. The programmed sequence of nucleotide addition
continues until the end of the program has been
reached.