|PyroMark Q24 Advanced||PyroMark Q24|
|Throughput||1–24 samples||1–24 samples|
|Running volume||25 µl||25 µl|
|PCR requirements||5–10 µl |
(~0.5–3 pmol of product)
|5–10 µl |
(~0.5–3 pmol of product)
|Read length (assay- and |
|10–140 bp or more||10–80 bp|
|Application software||PyroMark Q24 Advanced SW |
(requires firmware 1.5.6903 or higher)
|PyroMark Q24 SW 2.0|
|Software functionality||SEQ (de novo sequencing) |
|SQA (de novo sequencing) |
|Main applications||Complex mutation analysis |
Epigenetics (CpG and CpN analysis)
Resistance typing and microbial ID
|Mutation analysis |
|Compatible reagents||PyroMark Q24 Advanced Reagents |
PyroMark Q24 Advanced CpG Reagents
|PyroMark Q24 Gold Reagents|
|Sensitivity||2% mutation |
|2% mutation |
The new PyroMark Q24 Advanced Software (version 3.0.1) is compatible with Windows XP, Vista (32 and 64 bit), Windows 7 (32 and 64 bit), and Windows 10 (64 bit).
If you need to install the PyroMark Q24 software on a new computer replacing your old one, or after you reinstall or upgrade the operating system, you can reinstall the PyroMark Q24 software without purchasing a new license.
FAQ ID - 3473
For PyroMark Q24, PyroMark Q24 Advanced, PyroMark Q96 ID v2.5, and PyroMark CpG software, data are file-based. The original run files can be attached to email and sent to QIAGEN Technical Service.
For PyroMark Q96 ID 1.0, PyroMark Q96 MD/MDA 1.0, PSQ MA, and PSQ HS/HSA software, data are stored in an Oracle database and need exporting (See Instruction for run file export). Exported data can be attached to email and sent to QIAGEN Technical Service.
The log files from the PyroMark Q96 ID, PyroMark Q96 MD/MDA, PSQ MA, and PSQ HS/HSA instruments provide valuable information. See Instruction for log file retrieval) for log file retrieval instruction
Please send the log file along with the original run file to QIAGEN Technical Service.
Do not remove the USB stick while data are being copied. Corrupted data files are usually due to the interrupted copying process. Follow these steps to retrieve recently saved runs.
1. When the instrument is not processing, insert a USB stick into the USB port on the instrument.
2. Use the Up and Down buttons to select “Administration” in the menu and press OK.
3. Select “Copy Recently Saved Runs” and press OK.
4. Use the Up and Down buttons to select the run file(s) for retrieval and press Select.
5. When the instrument confirms that the run file(s) has been saved to the USB stick, press Close and remove the USB stick.
In addition, repeat the process with a different USB stick. If the issue persists, please contact QIAGEN Technical Service for further assistance.
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. Check the details at Managing PyroMark Instrument Methods.
The histogram is the theoretical peak pattern based on the Sequence to Analyze that you enter into the software, while the pyrogram is the experimental peak pattern that is detected by PyroMark instruments. The peak pattern in the pyrogram has to be in agreement with that in the histogram for a successful run. Overlaying the histogram on the pyrogram provides an invaluable tool for trouble-shooting Pyrosequencing runs. To do so, right click anywhere on the pyrogram and select Show Histogram on the pop-up menu.
Yes. The PyroMark Q24, PyroMark Q24 Advanced, and PyroMark Q96 v2.5 software provide multiple types of assay modes, which enable analyzing different types of samples at the same time.
A single peak height is the signal produced from a non-variable single nucleotide. The single peak height is an important criterion for trouble-shooting.
The optimal single peak height is 30 RLU (minimum 20 RLU) (Relative Light Unit) for PyroMark Q24 and PyroMark Q24 Advanced, 20 RLU (minimum 10 RLU) for PyroMark Q96 ID, and 100 RLU (minimum 50 RLU) for PyroMark Q96 MD instrument, respectively.
The PyroMark Q96 ID 1.0, PyroMark Q96 MD/MDA 1.0, PSQ MA, and PSQ HS/HSA software requires a functional Oracle database, which cannot be installed on an office computer. You can install the PyroMark software on an office computer and access the Oracle database on the PyroMark operator’s computer via network. The operator’s computer needs to be powered on while you are accessing the Oracle database remotely. Your IT department is responsible for the network setup.
This can result from very high signal in a well which results in cross-talk between neighbouring wells. It could also be that the PyroMark instrument's camera is misaligned (contact: QIAGEN Technical service).
Sodium hydroxide carry-over, too much template, and incorrectly stored reagent are the most common causes of wide peaks. Ensure correct volume of denaturation buffer is used on the vacuum prep station. Use 5–20 µl of PCR product depending on instrument models. Store reagent as described in the handbook.
If the issue persists, please send the run files (see Instruction for run file export) to QIAGEN Technical Service for further assistance.