PyroMark instruments dispense the correct volume of enzyme mix, substrate mix, and nucleotides using variable pulse times and dispensing pressures. The settings for pulse times and dispensing pressure are specific for each lot of PyroMark dispensing tips and cartridges and need to be updated in the PyroMark software being used to operate the instrument.
Dispensing pressures and pulse time settings must be checked and, if necessary, updated every time a cartridge from a new lot number is used. Reagent and nucleotide composition of a Pyrosequencing reaction is carefully balanced. When an incorrect pulse time or dispensing pressure is set, an incorrect volume of reagents and nucleotides will be dispensed into the reaction wells. This inaccurate dispensation can result in low or wide peaks or unexpected results in the Pyrogram. Other times, the effects may not be apparent in a Pyrogram or may result in failed Pyrosequencing runs.
To access detailed information for managing the instrument methods and pulse time settings for your specific Pyrosequencer, you must be logged in to the QIAGEN website and have a registered PyroMark instrument.