WAVE 系统低温标样谱图（ 56 0 C)
WAVE 系统 DNA 大小标样谱图（ 50 0 C)
DHPLC谱图疑难问题解答(WAVE系统低温标样和DNA大小标样为例）
Loss of Resolution on the Heteroduplexes, WAVE Low-Range Mutation Standard 
疑问解答：
- Oven not equilibrated before injection. Repeat injection to confirm results.
- Oven is running too hot. Loss of resolution between the first 2 peaks of the WAVE Low-Range Mutation Standard at 56° (heteroduplexes) indicates the oven needs calibration as long as resolution is maintained between the last 2 peaks (homoduplexes). Click here for instructions on Oven Calibration .
Loss of Resolution of the Homoduplexes, WAVE Low-Range Mutation Standard 
疑问解答：
- Oven not equilibrated before injection. Repeat injection to confirm results.
- Oven is running too cool. Loss of resolution between the last 2 peaks of the WAVE Low-Range Mutation Standard at 56° (homoduplexes) indicates the oven needs calibration as long as resolution is maintained between the first 2 peaks (heteroduplexes). Click here for instructions on Oven Calibration .
Loss of Resolution of both duplexes with poor peak shapes on WAVE Low-Range Mutation Standard
疑问解答：
- Separation Cartridge may need to be cleaned. Click here for instructions on Cartridge Cleaning .
- Has the Separation Cartridge been stored properly? Click here for information on Cartridge Care .
- Buffers may be old. Replace with Transgenomic WAVE? Optimized Buffers. Replace WAVE Optimized Buffers every two weeks. If you choose to make your own then Buffer A and Buffer B should be made every 5 days using proper reagents. See the Buffer Preparation Section.
- Inline filter may be clogged. Replace with new inline filter. (Click here for help with Changing the Inline Filter .)
- Lamp energy may be low. Check and replace as needed. (Click here for instructions on How to check and replace the lamp. )
Extremely Low Intensity for all peaks on WAVE Low-Range Mutation Standard 
疑问解答：
- Improper Volume? Be sure to inject at least 5μLs of the WAVE Low-Range Mutation Standard .
- Needle may not be going into the tube deep enough, and therefore may be unable to reach the sample.Or, the needle might be going too far into the vial and pressed against the bottom of the tube where it would be unable to take up the sample. Click here for instructions on checking and changing the needle depth.
- Air bubbles may be present that interfere with injection. Check vial to see if air bubbles are stuck to bottom of vial. Purge the pump to check for air bubbles in the flow path, and prime the syringe to check for air bubbles in the syringe. (See steps 5 and 6 of Basic Operation section for instructions.)
- Check lamp energy and sample/reference values. The lamp energy may be low or there may be an air bubble in the flow cell. If there is an air bubble in the flow cell, turn the flow rate up (1.0-1.5mL/min) for 10 minutes to force air bubble out. Click here for instructions on how to check lamp .
- Remove in-line filter from oven and replace with a union. Contaminated filters may bind DNA and inhibit detection. If intensity improves, replace union with a new inline filter. Replace filter once a month or every 1000 injections. Click here for information on how to install inline filters .
- Set up software to inject 75μLs of water. This can be done by doing one injection of 75μLs if the WAVE System has a 500μL (or larger) syringe, or by doing 3 injections of 25μLs each if the WAVE System has a 100 μL syringe. Fill a vial with 100μLs water, run the injection(s), and then measure the amount of water left in the vial. If it is not 25μLs, there may be a plug or leak somewhere along the needle or syringe flow path. Look at the injection port and see if any liquid is leaking from or sitting on top of the injection port. Click here for information on how to change an injection port seal .
No Injection Spike
疑问解答：
- This is what the chromatogram will look like if a 'blank', or zero-volume injection, was done.
- Check that the syringe volume and sample loop volume have been set correctly in the software. Typically, the syringe volume should be 500 and the sample loop volume should be 100, although this may vary from instrument to instrument.
- Is the injection valve turning? Turn autosampler off and then back on. The autosampler will run self-diagnostic tests. All is working well if all tests pass without an error message and the display shows the normal operation screen (see below) after initialization.
- See suggestions under the Low Intensity chromatogram.
Early Retention Times of WAVE Low-Range Mutation Standard
疑问解答：
- Have the buffers been made correctly? Tiny variations in acetonitrile concentration can greatly affect retention time. This is the cause of 90% of retention time issues. See Buffer Preparation section.
- If the system has not been allowed sufficient equilibration time, the retention times could shift. Always allow at least 3 minutes for equilibration when switching from a method that has a higher %B in it's 'Loading' gradient step. Always allow between 15 - 45 minutes equilibration after washing with 75% acetonitrile, depending on length of wash in the method.
- If air is in the flow path, retention times may shift. Purge the pump to remove air from flow path. Click here to learn how to purge the pump .
- Cartridges may start to show earlier retention times when they get old. If the cartridge has more than 4,000 injections, try a new one.
- Make sure that the oven is set to the correct temperature and that the software user has set no 'Time Shift' in the sample table.
Extra Peaks in the WAVE Sizing Control Sample
疑问解答：
- Extra peaks usually come from a degraded sample. Open a fresh standard, and try again.
- Air in the flow cell may appear as multiple peaks. Remove the cartridge, replace with a union, and turn the flow rate up to 1.5mL/min for 15 minutes to force air out of flow cell. Click here for help on using fittings, ferrules, and unions .
- A fracture in the cartridge bed may cause extra peaks. (This can occur if the cartridge is dropped or exposed to very high pressure.) Replace cartridge.
- Old or improperly made Syringe Wash solution may cause this type of performance. Replace syringe wash with Transgenomic WAVE? Optimized Syringe Wash or see Buffer Preparation .
Air Spike
Flat Line
疑问解答：
- Is the lamp on? Display screen will say Lamp Off if lamp is burned out. Click here for instructions on replacing the lamp.
- Is the cartridge hooked up to the correct detector and is the software configured for the detector in use?
- Is the cartridge installed?
|