QIAxcel Advanced System

Make sure the Gel Cartridge has been calibrated with the computer currently connected.  If not, please re-calibrate the Gel Cartridge.

Make sure the cartridge has equilibrated to room temperature (20°–25°C [68°– 77°F]) and stood upright in the cartridge stand for 20 min prior to use. Empty the buffer tray.

Wash the buffer tray in warm water and rinse thoroughly with deionized or reverse-osmosis water. Refill the buffer tray. Perform a control run with the alignment marker and bland samples (10 μl QX DNA Dilution Buffer). The marker bands should now appear within the chosen separation time.

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:

http://www.qiagen.com/products/catalog/automated-solutions/detection-and-analysis/qiaxcel-dna-kits#

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.

Unplug the QIAxcel or QIAxcel Advanced instrument and remove the fuse holder to verify that the fuses are good. If not, please replace the fuses (Cat No. 9241178 Fuse, 4A, 250VAC, QX). If this does not correct the problem, contact QIAGEN Technical Services.

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 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.

The revolutionary QIAxcel Advanced System replaces traditional, labor-intensive gel analysis of DNA and RNA.

• Rapid analysis of up to 96 samples without manual intervention
• Safety and convenience with ready-to-use gel cartridges
• Robust results for nucleic acid concentrations as low as 0.1 ng/ul
• Standardized and accurate analysis with a resolution down to 3-5 bp
• User-friendly analysis software that supports 21 CFR Part 11 compliance

See:  http://www.qiagen.com/Knowledge-and-Support/Videos-and-Virtual-Demos/Assay-Demos/QIAxcelAdvancedDemo/  for a virtual demonstration of the special software, features, and applications of the QIAxcel Advanced System.

The working alignment marker is an indication that the cartridge channel is working well. DNA samples that do not appear in all channels of the QIAxcel System or QIAxcel Advanced may be caused by:

1. Low sample volume (< 10 ul). The minimum required sample volume is 10 ul. 
2. Trapped air bubble in the well. The air bubble must be removed. 
3. High salt concentration in the sample. Dilute the sample with the QX DNA or RNA dilution buffer and rerun the sample.

Check that the lower and upper alignment markers are detected correctly, and that the threshold is set properly.

Verify that the Cartridge Purge Port label is removed. If it has been removed, verify that the alignment marker and samples are in the appropriate locations in the Buffer Tray and the 96-well Sample Tray used with the QIAxcel System or QIAxcel Advanced?.

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.

N2 cylinders on the QIAxcel Advanced should be changed when one or both low pressure warnings are shown in the "Status Information" panel.

The left panel of the screen in the "Process" environment is called the "Status Information" panel.  

"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.

The solutions used with the QIAxcel System or QIAxcel Advanced should not have to be changed for the life of the Gel Cartridge (based on the number of runs).  However, if any contamination is suspected, the Buffer Tray should be cleaned and the buffers should be changed immediately.

If the gel cartridge tips for the QIAxcel System and QIAxcel Advanced are in contact with air, the tips will dry out. There are 3 ways of preventing the tips from drying.

1. Store the cartridge in the instrument with the instrument in the “Park” position. The washing solution (8 ml) covered with mineral oil must be in the “Park” position of the Solution Tray. 
2. Store the cartridge in the Cartridge Stand with enough mineral oil in the well of the Cartridge Stand to cover the capillary tips. The Cartridge Stand should only be used for short term storage, i.e. when changing cartridges during one working day. 
3. Place the cartridge back into its packaging with the capillary tips gently inserted into the soft gel in the box.

The minimum sample volume required for the QIAxcel System and QIAxcel Advanced is 10 ul to guarantee sample injection in each channel.

If one of the capillaries of the gel cartridge for the QIAxcel System or QIAxcel Advanced is clogged, the corresponding lane in the gel-view image is typically black.

When running a QIAxcel Advanced instrument with the BioCalculator software, ensure to use version v3.2.07 or higher. You can download the latest version of the software from:

BioCalculator: http://www.qiagen.com/resources/Download.aspx?id=b68d8727-f0d7-44e5-acc5-2bb6ed50b63d&lang=en&ver=1

ScreenGel: http://www.qiagen.com/Products/Catalog/Automated-Solutions/Detection-and-Analysis/QIAxcel-ScreenGel-Software#resources

When running RNA samples on the QIAxcel System or QIAxcel Advanced, the standard RNA denaturation process has to be followed prior to running the sample.

• Add an equal volume of QX RNA Denaturation buffer to your RNA sample and/or QX RNA size Marker.
• Heat the solution at 70°C for 2 minutes on a heating block or in a PCR machine, then place on ice for 1 minute.
• Bring the total sample volume to 10 μl using QX RNA Dilution Buffer and mix the solution by gently pipetting up and down a few times.
• Analyze the samples immediately.

Yes, the QIAxcel System or QIAxcel Advanced can be connected to a local network.

The BioCalculator (version 3.2.05) and ScreenGel software have a feature to close the pressure valve after the software is idled for more than 30 minutes. To re-open the pressure valve, execute a command with the software, such as move the buffer tray, unlatch and latch the cartridge; alternatively, you can also exit the software and re-launch it. Also see FAQ ID 3001.

The sample DNA concentration might be too low. You can increase sample injection time.

A very strong sample in the same run can cause the threshold to be set too high. As a result, the signal from weaker samples is below this threshold. Try diluting the sample with high concentration with the QX dilution buffer.

High salt concentration in the samples, such as restriction enzyme digested DNA, can interfere with sample injection. Try reducing the salt concentration by diluting such samples with the QX dilution buffer.

If the issue persists, please send data files to QIAGEN Technical Services. See  <a href="/knowledge-and-support/faq/?ID=71818D37-21EF-40B0-BA30-6C8CC8CA82B9"><b>Where are the QIAxcel data stored</a></b>.

High or very low salt concentration could lead to the described appearance when running samples on the QIAxcel System and QIAxcel Advanced. Make sure QX DNA Dilution buffer has been used to dilute your samples. Dilute your samples 1:2 in QX DNA Dilution buffer and re-analyze.