Can I perform two 850 flow runs with an Ion 314 Chip?
You may, but W2 may run out in the middle of the second run. However, you can collect the data right before W2 runs out and reanalyze it after the run finishes. If you want complete data, please perform only one run per initialization.
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
How long can I wait until I start the run after initialization of my Ion PGM System?
If you are using the Ion PGM™ 200 Sequencing Kit v2, you can perform a run within 27 hours after initialization. If you are going to perform two runs, make sure the second one starts within 27 hours.
If you are using the Ion PGM™ 200 Sequencing Kit, the limit is shorter; you will need to perform a run within 10 hours.
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
What is the picture on the touchscreen during an Ion Proton run that looks like a bubble moving?
The touchscreen shows the voltage measured on the chip and there will be some contrast according the signal intensity which may look like bubbles.
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
What should I do if an error message "Chip reading inconsistent. Please replace chip and try again" appears during initialization of my Ion PGM System?
The pH may not have been properly measured or the volume of W3 may be too small. Please check the following:
1) That there is 50 mL of solution inside the W3 bottle
2) That the sippers and bottles attached to the instrument are not loose
3) That the color of the sippers on the instrument are gray and blue
If there is not enough W3, please add more until it reaches the 50 mL line on the bottle.
If you are accidentally using the dark blue sippers (for the dNTPs), the tip will not touch the solution, since it is too short. Replace the sippers, press the 'start' button, and if the same message appears, change the chip to a different one and try again.
If you get an error, shut down and restart the Ion PGM System. Discard the solution inside the W1 bottle, make new solution by adding 350 ul of 100 mM NaOH, and restart initialization.
Also, if you are using an Ion 316 Chip v2 on a v3.4 or older instrument, the instrument will not recognize the chip.
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
What should I do if the message "There may be a blockage or no NaOH in W1. Please check W1 and run line clear then try again" appears during initialization of my Ion PGM System?
Please check the following:
1) That there is solution left in the W1 bottle
2) That the sippers and bottles are not loose
If the sippers and bottles are not loose and there is at least 200 mL of solution in the W1 bottle, the line between W1 and W2 may be blocked. Please run the line clear.
If the same error message appears even after the line is clear, detach the reagent bottles from the Ion PGM System, water clean and shut down the instrument. Cap the reagent bottles. Restart the instrument and check the Ion PGM chip. Open the chip clamp and check that there is no solution standing in the port of the chip. If there is some solution, change the chip to another used chip or a new chip.
Place the W2 and W3 bottles back on the Ion PGM System. Discard the solution inside the W1 bottle, make more solution by adding 350 µL of 100 mM NaOH, and place it back on the Ion PGM System. Restart initialization.
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
