Cat. No. / ID: 929703
For analysis and typing of PCR products from different HLA loci using the QIAxcel Advanced and the Helmberg-SCORE software
For each cartridge used with the QIAxcel System, 5 additional intensity calibration runs have been programmed onto the smart key. If i.e. using a QIAxcel DNA High-resolution Cartridge, 105 will be displayed when using the cartridge for the first time – 100 sample runs, plus 5 intensity calibration runs.
The N2 bottle for the QIAxcel System should be changed when the Press1: in the Instruments Control Panel is “low”.
"Pressure 1" status: OK/Low. OK indicates adequate pressure for the sample run. LOW indicates that the N2 pressure is sufficient for the current sample run; however, the N2 cylinder should be replaced once the run has finished.
"Pressure 2" status: OK/Low. OK indicates adequate pressure for the sample run. LOW indicates that the N2 pressure is insufficient for the current sample run and the analysis will not be performed. The N2 cylinder should be replaced.
Refer to Appendix D of the QIAxcel DNA Handbook for hot water soaking and gel droplet testing. The handbook can be accessed by going to the Resources tab in the link below:
The intensity calibration procedure on the QIAxcel System and QIAxcel Advanced is performed to normalize the signal intensities across all 12 channels of the gel cartridge. This corrects for natural intensity reading variations between each capillary in the cartridge. A correction factor is determined and applied for every subsequent run performed with the cartridge.
The QIAxcel DNA Fast Analysis Cartridge can only be run on the QIAxcel Fast Analysis System with the BioCalculator Fast Analysis software and the grey software key.
All other QIAxcel Cartridges can only be run on the QIAxcel System with the BioCalculator software and the blue software key.
If lower and upper alignment markers used with the QIAxcel System or QIAxcel Advanced are not aligned correctly, open the channel of the sample that was incorrectly aligned. Check whether any unexpected extra peaks have been detected or if the alignment marker peaks do have any “shoulders” that are detected as separate peaks. Delete any extra peaks if necessary and re-analyze your data.
The concentration of the DNA marker, pUC18/HaeIII, provided in the QIAxcel DNA Kits is 5ug/ml in 1XTE buffer solution. Customers can run their own DNA size marker sample with different concentration levels using the provided Methods (i.e. M500, H500, L500 and so on).
The recommended method for all basic applications with the QIAxcel System is the M500 method. The BioCalculator software also allows customers to choose their own Sample Injection Time in the Sequence table. The M500 method provides 5KV for 20 seconds for the electro-kinetic injection of lower concentrated DNA samples. If the Injection Time is doubled or tripled (40 or 60 sec), you can expect double or triple intensities (also peak heights).
The Gel Cartridge does not need to be recalibrated when using a new alignment marker and/or DNA size marker with the QIAxcel System or QIAxcel Advanced. Calibration is required only once, i.e. when the first cartridge is run or when the cartridge is used on a different QIAxcel system than the one it has been calibrated on, or when using a different PC than the one used to calibrate initially.
When using this specific marker combination with the QIAxcel System or QIAxcel Advanced, i.e. for STR analysis, the 1.8 kb fragment lies outside of the range of detectable fragments and does not show up on the electropherogram and/or the gel view image.
Data for each cartridge intensity calibration on the QIAxcel System is stored in a single file named 'calibration.log'. This file is saved in the BioCalculator root directory C:\Program Files\BioCalculator.
If, for any reason, a different computer is used than the one containing the calibration.log file, the file should be transferred to the new computer. Otherwise, recalibration of the cartridge is required.
Likewise, if the QIAxcel Gel cartridge is used on a different instrument than the one it was calibrated on, another intensity calibration should be performed.
Check the QIAxcel computer “COM port” in the Windows Control Panel and then select the correct COM port number in the BioCalculator Instrument Control panel. Be sure to turn the Instrument on first prior to launching the BioCalculator software. If there is still “NO COM”, please contact QIAGEN Technical Services.